Insecticidal proteins from plants and methods for their use

ABSTRACT

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populations and for producing compositions with insecticidal activity.

CROSS REFERENCE

This application is a continuation of U.S. application Ser. No.16/980,087, filed Sep. 11, 2020 which is a national stage filing ofInternational application no. PCT/US19/21775, filed on Mar. 12, 2019which claims the benefit of US Provisional Application No. 62/642642filed Mar. 14, 2018, the disclosures of which are incorporated herein byreference in their entirety.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

The official copy of the sequence listing is submitted electronicallyvia EFS-Web as an xml file named “76751-373463.xml” created on Oct. 10,2022, and having a size of 932 kilobytes and is filed concurrently withthe specification. The sequence listing contained in this xml documentis part of the specification and is herein incorporated by reference inits entirety.

FIELD

This disclosure relates to the field of molecular biology. Provided arenovel genes that encode pesticidal proteins. These pesticidal proteinsand the nucleic acid sequences that encode them are useful in preparingpesticidal formulations and in the production of transgenicpest-resistant plants.

BACKGROUND

Biological control of insect pests of agricultural significance using amicrobial agent, such as fungi, bacteria or another species of insectaffords an environmentally friendly and a commercially attractivealternative to synthetic chemical pesticides. Generally speaking, theuse of biopesticides presents a lower risk of pollution andenvironmental hazards and biopesticides provide greater targetspecificity than is characteristic of traditional broad-spectrumchemical insecticides. In addition, biopesticides often cost less toproduce and thus improve economic yield for a wide variety of crops.

Certain species of microorganisms of the genus Bacillus are known topossess pesticidal activity against a range of insect pests includingLepidoptera, Diptera, Coleoptera, Hemiptera and others. Bacillusthuringiensis (Bt) and Bacillus popilliae are among the most successfulbiocontrol agents discovered to date. Insect pathogenicity has also beenattributed to strains of B. larvae, B. lentimorbus, B. sphaericus and B.cereus. Microbial insecticides, particularly those obtained fromBacillus strains, have played an important role in agriculture asalternatives to chemical pest control.

Crop plants have been developed with enhanced insect resistance bygenetically engineering crop plants to produce pesticidal proteins fromBacillus. For example, corn and cotton plants have been geneticallyengineered to produce pesticidal proteins isolated from strains ofBacillus thuringiensis. These genetically engineered crops are nowwidely used in agriculture and have provided the farmer with anenvironmentally friendly alternative to traditional insect-controlmethods. While they have proven to be very successful commercially,these genetically engineered, insect-resistant crop plants provideresistance to only a narrow range of the economically important insectpests. In some cases, insects can develop resistance to differentinsecticidal compounds, which raises the need to identify alternativebiological control agents for pest control.

Accordingly, there remains a need for new pesticidal proteins withincreased insecticidal activity, different spectrum of activity, and/ormode of action against insect pests, e.g., insecticidal proteins whichare active against a variety of insects in the order Lepidoptera and theorder Coleoptera including but not limited to insect pests that havedeveloped resistance to existing insecticides.

SUMMARY

In one aspect, compositions and methods for conferring pesticidalactivity to bacteria, plants, plant cells, tissues and seeds areprovided. Compositions include nucleic acid molecules encoding sequencesfor pesticidal and insecticidal polypeptides, vectors comprising thosenucleic acid molecules, and host cells comprising the vectors.Compositions also include the pesticidal polypeptide sequences andantibodies to those polypeptides. Compositions also comprise transformedbacteria, plants, plant cells, tissues and seeds.

In another aspect, isolated or recombinant nucleic acid molecules areprovided encoding IPD113 polypeptides including amino acidsubstitutions, deletions, insertions, and fragments thereof. Providedare isolated or recombinant nucleic acid molecules capable of encodingIPD113 polypeptides of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO:427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO:436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO:450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO:469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO:480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO:489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494, andSEQ ID NO: 495, as well as amino acid substitutions, deletions,insertions, fragments thereof, and combinations thereof. Nucleic acidsequences that are complementary to a nucleic acid sequence of theembodiments or that hybridize to a sequence of the embodiments are alsoencompassed. The nucleic acid sequences can be used in DNA constructs orexpression cassettes for transformation and expression in organisms,including microorganisms and plants. The nucleotide or amino acidsequences may be synthetic sequences that have been designed forexpression in an organism including, but not limited to, a microorganismor a plant.

In another aspect, IPD113 polypeptides are encompassed. Also providedare isolated or recombinant IPD113 polypeptides of SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO:420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO:430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO:440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO:453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO:473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO:483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO:492, SEQ ID NO: 494, and SEQ ID NO: 495, as well as amino acidsubstitutions, deletions, insertions, fragments thereof and combinationsthereof.

In another aspect, methods are provided for producing the polypeptidesand for using those polypeptides for controlling or killing aLepidopteran, Coleopteran, nematode, fungi, and/or Dipteran pests. Thetransgenic plants of the embodiments express one or more of thepesticidal sequences disclosed herein. In various embodiments, thetransgenic plant further comprises one or more additional genes forinsect resistance, for example, one or more additional genes forcontrolling Coleopteran, Lepidopteran, Hemipteran or nematode pests. Itwill be understood by one of skill in the art that the transgenic plantmay comprise any gene imparting an agronomic trait of interest.

In another aspect, methods for detecting the nucleic acids andpolypeptides of the embodiments in a sample are also included. A kit fordetecting the presence of an IPD113 polypeptide or detecting thepresence of a polynucleotide encoding an IPD113 polypeptide in a sampleis provided. The kit may be provided along with all reagents and controlsamples necessary for carrying out a method for detecting the intendedagent, as well as instructions for use.

In another aspect, the compositions and methods of the embodiments areuseful to produce organisms for the production of IPD113 polypeptidesand transgenic plants with enhanced pest resistance or tolerance. Theseorganisms and compositions comprising the organisms are desirable foragricultural purposes. The compositions of the embodiments are alsouseful for generating altered or improved proteins that have pesticidalactivity or for detecting the presence of IPD113 polypeptides.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A-1C shows a Phylogentic Tree, using the Neighbor Joining Methodin the ALIGNX® module of the Vector NTI® suite, of the IPD113 homologsof Table 3. FIG. 1C shows the IPD113Dh homolog family Phylogenic Treeand six closely related subgroups of the IPD113Dh homolog family membersare boxed and indicated with different line dashing.

FIG. 2A-2D shows an amino acid sequence alignment, using the ALIGNX®module of the Vector NTI® suite, of selected members of the IPD113homolog family: IPD113Aa (SEQ ID NO: 1); IPD113Ab (SEQ ID NO: 2);IPD113Bb (SEQ ID NO: 7); IPD113Bc (SEQ ID NO: 8); IPD113Db (SEQ ID NO:10); IPD113Dh (SEQ ID NO: 16); IPD113Ei (SEQ ID NO: 39); IPD113Ej (SEQID NO: 40); IPD113Fa (SEQ ID NO: 41); IPD113Fl (SEQ ID NO: 49); IPD113Gg(SEQ ID NO: 59); and IPD113Gh (SEQ ID NO: 60). The sequence diversity ishighlighted.

FIG. 3A-3C shows an amino acid sequence alignment, using the ALIGNX®module of the Vector NTI® suite, of the IPD113 homolog subgroup of:IPD113Aa (SEQ ID NO: 1); IPD113Ab (SEQ ID NO: 2); IPD113Ac (SEQ ID NO:3); IPD113Ad (SEQ ID NO: 4); IPD113Ae (SEQ ID NO: 5); IPD113Ba (SEQ IDNO: 6); and IPD113Bb (SEQ ID NO: 7). The sequence diversity ishighlighted. The two conserved cysteine (C) residues are indicated witha “▴” below the

IPD113Bb sequence (SEQ ID NO: 7).

FIG. 4A-4L shows an amino acid sequence alignment, using the ALIGNX®module of the Vector NTI® suite, of the IPD113Dh homolog subgroups ofFIG. 1C: IPD113Da (SEQ ID NO: 9); IPD113Db (SEQ ID NO: 10); IPD113Dc(SEQ ID NO: 11); IPD113Dd (SEQ ID NO: 12); IPD113De (SEQ ID NO: 13);IPD113Df (SEQ ID NO: 14); IPD113Dg (SEQ ID NO: 15); IPD113Dh (SEQ ID NO:16); IPD113Di (SEQ ID NO: 17); IPD113Dj (SEQ ID NO: 18); IPD113Dk (SEQID NO: 19); IPD113Dl (SEQ ID NO: 20); IPD113Dm (SEQ ID NO: 21); IPD113Dn(SEQ ID NO: 22); IPD113Do (SEQ ID NO: 23); IPD113Dp (SEQ ID NO: 24);IPD113Ds (SEQ ID NO: 27); IPD113Du (SEQ ID NO: 30); IPD113Ee (SEQ ID NO:35); IPD113Ef (SEQ ID NO: 36); IPD113Eg (SEQ ID NO: 37); IPD113Eh (SEQID NO: 38); IPD113Es (SEQ ID NO: 77); IPD113Dae (SEQ ID NO: 88);IPD113Daf (SEQ ID NO: 89); IPD113Dag (SEQ ID NO: 90); IPD113Dah (SEQ IDNO: 91); IPD113Eu (SEQ ID NO: 93); IPD113Ev (SEQ ID NO: 94); IPD113Ew(SEQ ID NO: 95); IPD113Ex (SEQ ID NO: 96); IPD113Dai (SEQ ID NO: 97);IPD113Daj (SEQ ID NO: 98); IPD113Dak (SEQ ID NO: 100); IPD113Dal (SEQ IDNO: 101); IPD113Dam (SEQ ID NO: 102); IPD113Ey (SEQ ID NO: 103);IPD113Ez (SEQ ID NO: 104); IPD113Eaa (SEQ ID NO: 105); IPD113Eab (SEQ IDNO: 106); IPD113Eac (SEQ ID NO: 107); IPD113Ead (SEQ ID NO: 110);IPD113Dan (SEQ ID NO: 111); IPD113Dao (SEQ ID NO: 112); IPD113Dap (SEQID NO: 113); and IPD113Daq (SEQ ID NO: 114). The sequence diversity ishighlighted. The five conserved cysteine (C) residues are indicated witha “▴” below the IPD113Dh sequence (SEQ ID NO: 16).

FIG. 5 shows the % leaf damage by CEW, ECB and FAW of individualtransgenic T0 maize events from constructs VECTOR 1 and VECTOR 2expressing genes encoding the IPD113Dh polypeptide (SEQ ID NO: 2)compared to the negative control events containing the construct lackinga IPD113Dh polynucleotide (Empty). Each “

” symbol represents an individual event.

DETAILED DESCRIPTION

It is to be understood that this disclosure is not limited to theparticular methodology, protocols, cell lines, genera, and reagentsdescribed, as such may vary. It is also to be understood that theterminology used herein is for describing particular embodiments only,and is not intended to limit the scope of the present disclosure.

As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the protein” includes reference to one or more proteinsand equivalents thereof. All technical and scientific terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which this disclosure belongs unless clearly indicatedotherwise.

The present disclosure is drawn to compositions and methods forcontrolling pests. The methods involve transforming organisms withnucleic acid sequences encoding IPD113 polypeptides. The nucleic acidsequences of the embodiments are useful for preparing plants andmicroorganisms that possess pesticidal activity. Thus, transformedbacteria, plants, plant cells, plant tissues and seeds are provided. Thecompositions include pesticidal nucleic acids and proteins of bacterialspecies. The nucleic acid sequences find use in the construction ofexpression vectors for subsequent transformation into organisms ofinterest, as probes for the isolation of other homologous (or partiallyhomologous) genes, and for the generation of altered IPD113 polypeptidesby methods, such as site directed mutagenesis, domain swapping or DNAshuffling. The IPD113 polypeptides find use in controlling or killingLepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematodepest populations and for producing compositions with pesticidalactivity. Insect pests of interest include, but are not limited to,Lepidoptera species including but not limited to: Corn Earworm, (CEW)(Helicoverpa zea); European Corn Borer (ECB) (Ostrinia nubialis),diamond-back moth, e.g., Helicoverpa zea Boddie; soybean looper, e.g.,Pseudoplusia includens Walker; and velvet bean caterpillar e.g.,Anticarsia gemmatalis Hübner.

By “pesticidal toxin” or “pesticidal protein” or “insecticidal protein”is used herein to refer to a toxin that has toxic activity against oneor more pests, including, but not limited to, members of theLepidoptera, Diptera, Hemiptera and Coleoptera orders or the Nematodaphylum or a protein that has homology to such a protein. Pesticidalproteins have been isolated from organisms including, for example,Bacillus sp., Pseudomonas sp., Photorhabdus sp., Xenorhabdus sp.,Clostridium bifermentans and Paenibacillus popilliae. Pesticidalproteins include but are not limited to: insecticidal proteins fromPseudomonas sp. such as PSEEN3174 (Monalysin; (2011) PLoS Pathogens7:1-13); from Pseudomonas protegens strain CHA0 and Pf-5 (previouslyfluorescens) (Pechy-Tarr, (2008) Environmental Microbiology10:2368-2386; GenBank Accession No. EU400157); from Pseudomonastaiwanensis (Liu, et al., (2010) J. Agric. Food Chem., 58:12343-12349)and from Pseudomonas pseudoalcaligenes (Zhang, et al., (2009) Annals ofMicrobiology 59:45-50 and Li, et al., (2007) Plant Cell Tiss. OrganCult. 89:159-168); insecticidal proteins from Photorhabdus sp. andXenorhabdus sp. (Hinchliffe, et al., (2010) The Open Toxicology Journal,3:101-118 and Morgan, et al., (2001) Applied and Envir. Micro.67:2062-2069); U.S. Pat. Nos. 6,048,838, and 6,379,946; a PIP-1polypeptide of U.S. Pat. No. 9,688,730; an AfIP-1A and/or AfIP-1Bpolypeptide of U.S. Pat. No. 9,475,847; a PIP-47 polypeptide of USPublication Number US20160186204; an IPD045 polypeptide, an IPD064polypeptide, an IPD074 polypeptide, an IPD075 polypeptide, and an IPD077polypeptide of PCT Publication Number WO 2016/114973; an IPD080polypeptide of PCT Serial Number PCT/US17/56517; an IPD078 polypeptide,an IPD084 polypeptide, an IPD085 polypeptide, an IPD086 polypeptide, anIPD087 polypeptide, an IPD088 polypeptide, and an IPD089 polypeptide ofSerial Number PCT/US17/54160; PIP-72 polypeptide of US PatentPublication Number US20160366891; a PtIP-50 polypeptide and a PtIP-65polypeptide of US Publication Number US20170166921; an IPD098polypeptide, an IPD059 polypeptide, an IPD108 polypeptide, an IPD109polypeptide of US Serial number 62/521084; a PtIP-83 polypeptide of USPublication Number US20160347799; a PtIP-96 polypeptide of USPublication Number US20170233440; an IPD079 polypeptide of PCTPublication Number WO2017/23486; an IPD082 polypeptide of PCTPublication Number WO 2017/105987, an IPD090 polypeptide of SerialNumber PCT/US17/30602, an IPD093 polypeptide of U.S. Ser. No. 62/434020;an IPD103 polypeptide of Serial Number PCT/US17/39376; an IPD101polypeptide of U.S. Ser. No. 62/438179; an IPD121 polypeptide of USSerial Number U.S. 62/508,514; and δ-endotoxins including, but notlimited to a Cry1, Cry2, Cry3, Cry4, Cry5, Cry6, Cry7, Cry8, Cry9,Cry10, Cry11, Cry12, Cry13, Cry14, Cry15, Cry16, Cry17, Cry18, Cry19,Cry20, Cry21, Cry22, Cry23, Cry24, Cry25, Cry26, Cry27, Cry28, Cry29,Cry30, Cry31, Cry32, Cry33, Cry34, Cry35,Cry36, Cry37, Cry38, Cry39,Cry40, Cry41, Cry42, Cry43, Cry44, Cry45, Cry46, Cry47, Cry49, Cry50,Cry51, Cry52, Cry53, Cry54, Cry55, Cry56, Cry57, Cry58, Cry59, Cry60,Cry61, Cry62, Cry63, Cry64, Cry65, Cry66, Cry67, Cry68, Cry69, Cry70,Cry71, and Cry 72 classes of δ-endotoxin polypeptides and the B.thuringiensis cytolytic cyt1 and cyt2 genes. Members of these classes ofB. thuringiensis insecticidal proteins (see, Crickmore, et al.,“Bacillus thuringiensis toxin nomenclature” (2011), atlifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/ which can be accessed onthe world-wide web using the “www” prefix).

Examples of δ-endotoxins also include but are not limited to Cry1Aproteins of U.S. Pat. Nos. 5,880,275, 7,858,849, and 8,878,007; a Cry1Acmutant of U.S. Pat. No. 9,512,187; a DIG-3 or DIG-11 toxin (N-terminaldeletion of α-helix 1 and/or α-helix 2 variants of cry proteins such asCry1A, Cry3A) of U.S. Pat. Nos. 8,304,604, 8.304,605 and 8,476,226;Cry1B of U.S. patent application Ser. No. 10/525,318, US PatentApplication Publication Number US20160194364, and U.S. Pat. Nos.9,404,121 and 8,772,577; Cry1B variants of PCT Publication NumberWO2016/61197 and Serial Number PCT/US17/27160; Cry1C of U.S. Pat. No.6,033,874; Cry1D protein of US20170233759; a Cry1E protein of PCT SerialNumber PCT/US17/53178; a Cry1F protein of U.S. Pat. Nos. 5,188,960 and6,218,188; Cry1A/F chimeras of U.S. Pat. Nos. 7,070,982; 6,962,705 and6,713,063; a Cry1I protein of PCT Publication number WO 2017/0233759; aCry1J variant of US Publication US20170240603; a Cry2 protein such asCry2Ab protein of U.S. Pat. No. 7,064,249 and Cry2A.127 protein of U.S.Pat. No. 7208474; a Cry3A protein including but not limited to anengineered hybrid insecticidal protein (eHIP) created by fusing uniquecombinations of variable regions and conserved blocks of at least twodifferent Cry proteins (US Patent Application Publication Number2010/0017914); a Cry4 protein; a Cry5 protein; a Cry6 protein; Cry8proteins of U.S. Pat. Nos. 7,329,736, 7,449,552, 7,803,943, 7,476,781,7,105,332, 7,339,092, 7,378,499, 7,462,760, and 9,593,345; a Cry9protein such as such as members of the Cry9A, Cry9B, Cry9C, Cry9D, Cry9Eand Cry9F families including the Cry9 protein of U.S. Pat. Nos.9,000,261 and 8,802,933, and US Serial Number WO 2017/132188; a Cry15protein of Naimov, et al., (2008) Applied and EnvironmentalMicrobiology, 74:7145-7151; a Cry14 protein of U.S. Pat. No. 8,933,299;a Cry22, a Cry34Ab1 protein of U.S. Pat. No. 6,127,180, 6,624,145 and6,340,593; a truncated Cry34 protein of U.S. Pat. No. 8,816,157; aCryET33 and cryET34 protein of U.S. Pat. Nos. 6,248,535, 6,326,351,6,399,330, 6,949,626, 7,385,107 and 7,504,229; a CryET33 and CryET34homologs of US Patent Publication Number 2006/0191034, 2012/0278954, andPCT Publication Number WO 2012/139004; a Cry35Ab1 protein of U.S. Pat.Nos. 6,083,499, 6,548,291 and 6,340,593; a Cry46 protein of U.S. Pat.No. 9,403,881, a Cry 51 protein, a Cry binary toxin; a TIC901 or relatedtoxin; TIC807 of US Patent Application Publication Number 2008/0295207;TIC853 of U.S. Pat. No. 8,513,493; ET29, ET37, TIC809, TIC810, TIC812,TIC127, TIC128 of PCT US 2006/033867; engineered Hemipteran toxicproteins of US Patent Application Publication Number US20160150795,AXMI-027, AXMI-036, and AXMI-038 of U.S. Pat. No. 8,236,757; AXMI-031,AXMI-039, AXMI-040, AXMI-049 of U.S. Pat. No. 7,923,602; AXMI-018,AXMI-020 and AXMI-021 of WO 2006/083891; AXMI-010 of WO 2005/038032;AXMI-003 of WO 2005/021585; AXMI-008 of US Patent ApplicationPublication Number 2004/0250311; AXMI-006 of US Patent ApplicationPublication Number 2004/0216186; AXMI-007 of US Patent ApplicationPublication Number 2004/0210965; AXMI-009 of US Patent ApplicationNumber 2004/0210964; AXMI-014 of US Patent Application PublicationNumber 2004/0197917; AXMI-004 of US Patent Application PublicationNumber 2004/0197916; AXMI-028 and AXMI-029 of WO 2006/119457; AXMI-007,AXMI-008, AXMI-0080rf2, AXMI-009, AXMI-014 and AXMI-004 of WO2004/074462; AXMI-150 of U.S. Pat. No. 8,084,416; AXMI-205 of US PatentApplication Publication Number 2011/0023184; AXMI-011, AXMI-012,AXMI-013, AXMI-015, AXMI-019, AXMI-044, AXMI-037, AXMI-043, AXMI-033,AXMI-034, AXMI-022, AXMI-023, AXMI-041, AXMI-063 and AXMI-064 of USPatent Application Publication Number 2011/0263488; AXMI046, AXMI048,AXMI050, AXMI051, AXMI052, AXMI053, AXMI054, AXMI055, AXMI056, AXMI057,AXMI058, AXMI059, AXMI060, AXMI061, AXMI067, AXMI069, AXMI071, AXMI072,AXMI073, AXMI074, AXMI075, AXMI087, AXMI088, AXMI093, AXMI070, AXMI080,AXMI081, AXMI082, AXMI091, AXMI092, AXMI096, AXMI097, AXMI098, AXMI099,AXMI100, AXMI101, AXMI102, AXMI103, AXMI104, AXMI107, AXMI108, AXMI109,AXMI110, AXMI111, AXMI112, AXMI114, AXMI116, AXMI117, AXMI118, AXMI119,AXMI120, AXMI121, AXMI122, AXMI123, AXMI124, AXMI125, AXMI126, AXMI127,AXMI129, AXMI151, AXMI161, AXMI164, AXMI183, AXMI132, AXMI137, AXMI138of U.S. Pat. Nos. 8,461,421 and 8,461,422; AXMI-R1 and related proteinsof US Patent Application Publication Number 2010/0197592; AXMI221Z,AXMI222z, AXMI223z, AXMI224z and AXMI225z of WO 2011/103248; AXMI218,AXMI219, AXMI220, AXMI226, AXMI227, AXMI228, AXMI229, AXMI230 andAXMI231 of WO 2011/103247; AXMI-115, AXMI-113, AXMI-005, AXMI-163 andAXMI-184 of U.S. Pat. No. 8,334,431; AXMI-001, AXMI-002, AXMI-030,AXMI-035 and AXMI-045 of US Patent Application Publication Number2010/0298211; AXMI-066 and AXMI-076 of US Patent Application PublicationNumber 2009/0144852; AXMI128, AXMI130, AXMI131, AXMI133, AXMI140,AXMI141, AXMI142, AXMI143, AXMI144, AXMI146, AXMI148, AXMI149, AXMI152,AXMI153, AXMI154, AXMI155, AXMI156, AXMI157, AXMI158, AXMI162, AXMI165,AXMI166, AXMI167, AXMI168, AXMI169, AXMI170, AXMI171, AXMI172, AXMI173,AXMI174, AXMI175, AXMI176, AXMI177, AXMI178, AXMI179, AXMI180, AXMI181,AXMI182, AXMI185, AXMI186, AXMI187, AXMI188, AXMI189 of U.S. Pat. No.8,318,900; AXMI079, AXMI080, AXMI081, AXMI082, AXMI091, AXMI092,AXMI096, AXMI097, AXMI098, AXMI099, AXMI100, AXMI101, AXMI102, AXMI103,AXMI104, AXMI107, AXMI108, AXMI109, AXMI110, dsAXMI111, AXMI112,AXMI114, AXMI116, AXMI117, AXMI118, AXMI119, AXMI120, AXMI121, AXMI122,AXMI123, AXMI124, AXMI1257, AXMI1268, AXMI127, AXMI129, AXMI164,AXMI151, AXMI161, AXMI183, AXMI132, AXMI138, AXMI137 of U.S. Pat. No.8,461,421; AXMI192 of U.S. Pat. No. 8,461,415; AXMI281 of US PatentApplication Publication Number US20160177332; AXMI422 of U.S. Pat. Nos.8,252,872; cry proteins such as Cry1A and Cry3A having modifiedproteolytic sites of U.S. Pat. No. 8,319,019; a Cry1Ac, Cry2Aa andCry1Ca toxin protein from Bacillus thuringiensis strain VBTS 2528 of USPatent Application Publication Number 2011/0064710. The Cry proteinsMP032, MP049, MP051, MP066, MP068, MP070, MP091S, MP109S, MP114, MP121,MP134S, MP183S, MP185S, MP186S, MP195S, MP197S, MP208S, MP209S, MP212S,MP214S, MP217S, MP222S, MP234S, MP235S, MP237S, MP242S, MP243, MP248,MP249S, MP251 M, MP252S, MP253, MP259S, MP287S, MP288S, MP295S, MP296S,MP297S, MP300S, MP304S, MP306S, MP310S, MP312S, MP314S, MP319S, MP325S,MP326S, MP327S, MP328S, MP334S, MP337S, MP342S, MP349S, MP356S, MP359S,MP360S, MP437S, MP451S, MP452S, MP466S, MP468S, MP476S, MP482S, MP522S,MP529S, MP548S, MP552S, MP562S, MP564S, MP566S, MP567S, MP569S, MP573S,MP574S, MP575S, MP581S, MP590, MP594S, MP596S, MP597, MP599S, MP600S,MP601S, MP602S, MP604S, MP626S, MP629S, MP630S, MP631S, MP632S, MP633S,MP634S, MP635S, MP639S, MP640S, MP644S, MP649S, MP651S, MP652S, MP653S,MP661S, MP666S, MP672S, MP696S, MP704S, MP724S, MP729S, MP739S, MP755S,MP773S, MP799S, MP800S, MP801S, MP802S, MP803S, MP805S, MP809S, MP815S,MP828S, MP831S, MP844S, MP852, MP865S, MP879S, MP887S, MP891S, MP896S,MP898S, MP935S, MP968, MP989, MP993, MP997, MP1049, MP1066, MP1067,MP1080, MP1081, MP1200, MP1206, MP1233, and MP1311 of U.S. Ser. No.62/607372. The insecticidal activity of Cry proteins is well known toone skilled in the art (for review, see, van Frannkenhuyzen, (2009) J.Invert. Path. 101:1-16). The use of Cry proteins as transgenic planttraits and Cry-transgenic plants including but not limited to plantsexpressing Cry1Ac, Cry1Ac+Cry2Ab, Cry1Ab, Cry1A.105, Cry1F, Cry1Fa2,Cry1F+Cry1Ac, Cry2Ab, Cry3A, mCry3A, Cry3Bb1, Cry34Ab1, Cry35Ab1, Vip3A,mCry3A, Cry9c and CBI-Bt have received regulatory approval (see,Sanahuja, (2011) Plant Biotech Journal 9:283-300 and the CERA. (2010) GMCrop Database Center for Environmental Risk Assessment (CERA), ILSIResearch Foundation, Washington D.C. atcera-gmc.org/index.php?action=gm_crop_database which can be accessed onthe world-wide web using the “www” prefix). More than one pesticidalproteins can also be expressed in plants such as Vip3Ab & Cry1Fa(US2012/0317682); Cry1BE & Cry1F (US2012/0311746); Cry1CA & Cry1AB(US2012/0311745); Cry1F & CryCa (US2012/0317681); Cry1DA & Cry1BE(US2012/0331590); Cry1DA & Cry1Fa (US2012/0331589); Cry1AB & Cry1BE(US2012/0324606); Cry1Fa & Cry2Aa and Cry1I & Cry1E (US2012/0324605);Cry34Ab/35Ab & Cry6Aa (US20130167269); Cry34Ab/VCry35Ab & Cry3Aa(US20130167268); Cry1Da & Cry1Ca (US 9796982); Cry3Aa & Cry6Aa (US9798963); and Cry3A & Cry1Ab or Vip3Aa (U.S. Pat. No. 9,045,766).Pesticidal proteins also include insecticidal lipases including lipidacyl hydrolases of U.S. Pat. No. 7,491,869, and cholesterol oxidasessuch as from Streptomyces (Purcell et al. (1993) Biochem Biophys ResCommun 15:1406-1413). Pesticidal proteins also include VIP (vegetativeinsecticidal proteins) toxins of U.S. Pat. Nos. 5,877,012, 6,107,2796,137,033, 7,244,820, 7,615,686, and 8,237,020 and the like. Other VIPproteins (see, lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/vip.html)which can be accessed on the world-wide web using the “www” prefix).Pesticidal proteins also include Cyt proteins including Cyt1A variantsof PCT Serial Number PCT/US2017/000510; Pesticidal proteins also includetoxin complex (TC) proteins, obtainable from organisms such asXenorhabdus, Photorhabdus and Paenibacillus (see, U.S. Pat. Nos.7,491,698 and 8,084,418). Some TC proteins have “stand alone”insecticidal activity and other TC proteins enhance the activity of thestand-alone toxins produced by the same given organism. The toxicity ofa “stand-alone” TC protein (from Photorhabdus, Xenorhabdus orPaenibacillus, for example) can be enhanced by one or more TC protein“potentiators” derived from a source organism of a different genus.There are three main types of TC proteins. As referred to herein, ClassA proteins (“Protein A”) are stand-alone toxins. Class B proteins(“Protein B”) and Class C proteins (“Protein C”) enhance the toxicity ofClass A proteins. Examples of Class A proteins are TcbA, TcdA, XptA1 andXptA2. Examples of Class B proteins are TcaC, TcdB, XptB1Xb and XptC1Wi.Examples of Class C proteins are TccC, XptC1Xb and XptB1Wi. Pesticidalproteins also include spider, snake and scorpion venom proteins.Examples of spider venom peptides include but not limited to lycotoxin-1peptides and mutants thereof (U.S. Pat. No. 8,334,366). The combinationsgenerated can also include multiple copies of any one of thepolynucleotides of interest.

In some embodiments, the IPD113 polypeptide includes an amino acidsequence deduced from the full-length nucleic acid sequence disclosedherein and amino acid sequences that are shorter than the full-lengthsequences, either due to the use of an alternate downstream start siteor due to processing that produces a shorter protein having pesticidalactivity. Processing may occur in the organism the protein is expressedin or in the pest after ingestion of the protein.

Thus, provided herein are novel isolated or recombinant nucleic acidsequences that confer pesticidal activity. Also provided are the aminoacid sequences of IPD113 polypeptides. The protein resulting fromtranslation of these IPD113 genes allows cells to control or killcertain pests that ingest it.

IPD113 Proteins and Variants and Fragments Thereof

IPD113 polypeptides are encompassed by the disclosure. “IPD113polypeptide” and “IPD113 protein” as used herein interchangeably refersto a polypeptide having insecticidal activity including but not limitedto insecticidal activity against one or more insect pests of theLepidoptera order, and is sufficiently homologous to the IPD113Dhpolypeptide of SEQ ID NO: 16. A variety of IPD113 polypeptides arecontemplated. Sources of IPD113 polypeptides or related proteins includefern or other primitive plant species selected from, but not limited to,the Genus Pteris, Polypodium, Nephrolepis, Colysis, Tectaria, Davallia,Polystichum, Adiantum, Asplenium, Blechnum, Lygodium, Ophioglossum,Pyrrosia, Doryopteris, Dryopteris, Pellaea, Gymnocarpium, Cheilanthes,Pteridium, Christella, Lastreopsis, Campyloneurum, Hemionitis,Selliguea, and Arachniodes.

In some embodiments, the IPD113 polypeptide is derived from a species inthe Genus Pteris. In some embodiments, the IPD113 polypeptide is derivedfrom a Pteris species selected from but not limited to Pteris aberrans,Pteris abyssinica, Pteris actiniopteroides, Pteris adscensionis, Pterisalbersii, Pteris albertiae, Pteris altissima, Pteris amoena, Pterisangustata, Pteris angustipinna, Pteris angustipinnula, Pterisappendiculifera, Pteris arborea, Pteris argyraea, Pteris aspericaulis,Pteris asperula, Pteris atrovirens, Pteris auquieri, Pterisaustrosinica, Pteris bahamensis, Pteris bakeri, Pteris baksaensis,Pteris balansae, Pteris bambusoides, Pteris barbigera, Pterisbarombiensis, Pteris bavazzanoi, Pteris beecheyana, Pteris bella, Pterisberteroana, Pteris biaurita, Pteris biformis, Pteris blanchetiana,Pteris blumeana, Pteris boninensis, Pteris brassii, Pteris brevis,Pteris brooksiana, Pteris buchananii, Pteris buchtienii, Pterisburtonii, Pteris cadieri, Pteris caesia, Pteris caiyangheensis, Pteriscalcarea, Pteris calocarpa, Pteris catoptera, Pteris chiapensis, Pterischilensis, Pteris christensenii, Pteris chrysodioides, Pteris ciliaris,Pteris clemensiae, Pteris comans, Pteris commutata, Pteris concinna,Pteris confertinervia, Pteris confusa, Pteris congesta, Pterisconsanguinea, Pteris coriacea, Pteris crassiuscula, Pteris cretica,Pteris croesus, Pteris cryptogrammoides, Pteris cumingii, Pterisdactylina, Pteris daguensis, Pteris dalhousiae, Pteris dataensis, Pterisdayakorum, Pteris decrescens, Pteris decurrens, Pteris deflexa, Pterisdeltea, Pteris deltodon, Pteris deltoidea, Pteris dentata, Pterisdenticulata, Pteris dispar, Pteris dissimilis, Pteris dissitifolia,Pteris distans, Pteris droogmaniana, Pteris edanyoi, Pteris ekmanii,Pteris elmeri, Pteris elongatiloba, Pteris endoneura, Pteris ensiformis,Pteris esquirolii, Pteris excelsa, Pteris famatinensis, Pteris fauriei,Pteris finotii, Pteris flava, Pteris formosana, Pteris fraseri, Pterisfriesii, Pteris gallinopes, Pteris geminata, Pteris gigantea, Pterisglaucovirens, Pteris goeldii, Pteris gongalensis, Pteris grandifolia,Pteris grevilleana, Pteris griffithii, Pteris griseoviridis, Pterisguangdongensis, Pteris guizhouensis, Pteris haenkeana, Pteris hamulosa,Pteris hartiana, Pteris heteroclita, Pteris heteromorpha, Pterisheterophlebia, Pteris hillebrandii, Pteris hirsutissima, Pteris hirtula,Pteris hispaniolica, Pteris holttumii, Pteris hondurensis, Pterishookeriana, Pteris hossei, Pteris hostmanniana, Pteris hui, Pterishumbertii, Pteris hunanensis, Pteris inaequalis, Pteris incompleta,Pteris inermis, Pteris insigni, Pteris intricata, Pteris intromissa,Pteris irregularis, Pteris iuzonensis, Pteris izuensis, Pterisjohannis-winkleri, Pteris junghuhnii, Pteris kawabatae, Pteris keysseri,Pteris khasiana, Pteris kidoi, Pteris kinabaluensis, Pteris kingiana,Pteris kiuschiuensis, Pteris laevis, Pteris lanceifolia, Pteris lastii,Pteris laurea, Pteris laurisilvicola, Pteris lechleri, Pterislepidopoda, Pteris leptophylla, Pteris liboensis, Pteris ligulata,Pteris limae, Pteris linearis, Pteris litoralis, Pteris livida, Pterisloheri, Pteris longifolia, Pteris longipes, Pteris longipetiolulata,Pteris longipinna, Pteris longipinnula, Pteris luederwaldtii, Pterisluschnathiana, Pteris luzonensis, Pteris lydgatei, Pteris macgregorii,Pteris macilenta, Pteris maclurei, Pteris maclurioides, Pterismacracantha, Pteris macrodon, Pteris macrophylla, Pteris macroptera,Pteris madagascarica, Pteris majestica, Pteris malipoensis, Pterismanniana, Pteris melanocaulon, Pteris melanorhachis, Pteris menglaensis,Pteris mertensioides, Pteris mettenii, Pteris micracantha, Pterismicrodictyon, Pteris microlepis, Pteris microptera, Pteris mildbraedii,Pteris moluccana, Pteris monghaiensis, Pteris montis-wilhelminae, Pterismorii, Pteris mucronulata, Pteris multiaurita, Pteris multifida, Pterismuricata, Pteris muricatopedata, Pteris muricella, Pteris mutilata,Pteris natiensis, Pteris navarrensis, Pteris nipponica, Pterisnovae-caledoniae, Pteris obtusiloba, Pteris occidentalisinica, Pterisolivacea, Pteris opaca, Pteris oppositipinnata, Pteris orientalis,Pteris orizabae, Pteris oshimensis, Pteris otaria, Pteris pachysora,Pteris pacifica, Pteris paleacea, Pteris papuana, Pteris parhamii,Pteris paucinervata, Pteris paucipinnata, Pteris paulistana, Pterispearcei, Pteris pedicellata, Pteris pediformis, Pteris pellucida, Pterisperrieriana, Pteris perrottetii, Pteris philippinensis, Pterisphuluangensis, Pteris pilosiuscula, Pteris plumbea, Pteris pluricaudata,Pteris podophylla, Pteris polita, Pteris polyphylla, Pterisporphyrophlebia, Pteris praetermissa, Pteris preussii, Pteris prolifera,Pteris propinqua, Pteris pseudolonchitis, Pteris pseudopellucida, Pterispteridioides, Pteris puberula, Pteris pulchra, Pteris pungens, Pterispurdoniana, Pteris purpureorachis, Pteris quadriaurita, Pterisquinquefoliata, Pteris quinquepartita, Pteris radicans, Pteris ramosii,Pteris rangiferina, Pteris reducta, Pteris remotifolia, Pteris reptans,Pteris rigidula, Pteris rosenstockii, Pteris roseo-lilacina, Pterisryukyuensis, Pteris satsumana, Pteris saxatilis, Pteris scabra, Pterisscabripes, Pteris schlechteri, Pteris schwackeana, Pteris semiadnata,Pteris semipinnata, Pteris sericea, Pteris setigera, Pterissetuloso-costulata, Pteris shimenensis, Pteris shimianensis, Pterissilvatica, Pteris similis, Pteris simplex, Pteris sintenensis, Pterisspeciosa, Pteris splendens, Pteris splendida, Pteris squamaestipes,Pteris squamipes, Pteris stenophylla, Pteris stridens, Pterisstriphnophylla, Pteris subindivisa, Pteris subquinata, Pterissubsimplex, Pteris sumatrana, Pteris swartziana, Pteris taiwanensis,Pteris talamauana, Pteris tapeinidiifolia, Pteris tarandus, Pteristenuissima, Pteris togoensis, Pteris torricelliana, Pteris trachyrachis,Pteris transparens, Pteris tremula, Pteris treubii, Pteris tricolor,Pteris tripartita, Pteris tussaci, Pteris umbrosa, Pteris undulatipinna,Pteris usambarensis, Pteris vaupelii, Pteris venusta, Pterisverticillata, Pteris vieillardii, Pteris viridissima, Pteris vitiensis,Pteris vittata, Pteris wallichiana, Pteris wangiana, Pteris warburgii,Pteris werneri, Pteris whitfordii, Pteris woodwardioides, Pteriswulaiensis, Pteris yakuinsularis, Pteris yamatensis, Pteriszahlbruckneriana, and Pteris zippelii.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPolypodium L. In some embodiments, the IPD113 polypeptide is derivedfrom a fern species in the Order Polypodiales, Family Polypodiaceae,Genus Polypodium L. selected from, but not limited to, Polypodiumabsidatum, Polypodium acutifolium, Polypodium adiantiforme, Polypodiumaequale, Polypodium affine, Polypodium albidopaleatum, Polypodiumalcicorne, Polypodium alfarii, Polypodium alfredii, Polypodium alfrediivar. curtii, Polypodium allosuroides, Polypodium alsophilicola,Polypodium amamianum, Polypodium amoenum, Polypodium amorphum,Polypodium anetioides, Polypodium anfractuosum, Polypodium anguinum,Polypodium angustifolium f. remotifolia, Polypodium angustifolium var.amphostenon, Polypodium angustifolium var. heterolepis, Polypodiumangustifolium var. monstrosa, Polypodium angustipaleatum, Polypodiumangustissimum, Polypodium anisomeron var. pectinatum, Polypodiumantioquianum, Polypodium aoristisorum, Polypodium apagolepis, Polypodiumapicidens, Polypodium apiculatum, Polypodium apoense, Polypodiumappalachianum, Polypodium appressum, Polypodium arenarium, Polypodiumargentinum, Polypodium argutum, Polypodium armatum, Polypodiumaromaticum, Polypodium aspersum, Polypodium assurgens, Polypodium atrum,Polypodium auriculatum, Polypodium balaonense, Polypodium balliviani,Polypodium bamleri, Polypodium bangii, Polypodium bartlettii, Polypodiumbasale, Polypodium bemoullii, Polypodium biauritum, Polypodium bifrons,Polypodium blepharodes, Polypodium bolivari, Polypodium bolivianum,Polypodium bolobense, Polypodium bombycinum, Polypodium bombycinum var.insularum, Polypodium bradeorum, Polypodium bryophilum, Polypodiumbryopodum, Polypodium buchtienii, Polypodium buesii, Polypodiumbulbotrichum, Polypodium caceresii, Polypodium californicum f.brauscombii, Polypodium californicum f. parsonsiae, Polypodiumcalifornicum, Polypodium calophlebium, Polypodium calvum, Polypodiumcamptophyllarium var. abbreviatum, Polypodium capitellatum, Polypodiumcarpinterae, Polypodium chachapoyense, Polypodium chartaceum, Polypodiumchimantense, Polypodium chiricanum, Polypodium choquetangense,Polypodium christensenii, Polypodium christii, Polypodium chrysotrichum,Polypodium ciliolepis, Polypodium cinerascens, Polypodium collinsii,Polypodium colysoides, Polypodium confluens, Polypodium conforme,Polypodium confusum, Polypodium congregatifolium, Polypodium connellii,Polypodium consimile var. bourgaeanum, Polypodium consimile var. minor,Polypodium conterminans, Polypodium contiguum, Polypodium cookii,Polypodium coriaceum, Polypodium coronans, Polypodium costaricense,Polypodium costatum, Polypodium crassifolium f. angustissimum,Polypodium crassifolium var. longipes, Polypodium crassulum, Polypodiumcraterisorum, Polypodium cryptum, Polypodium crystalloneuron, Polypodiumcucullatum var. planum, Polypodium cuencanum, Polypodium cumingianum,Polypodium cupreolepis, Polypodium curranii, Polypodium curvans,Polypodium cyathicola, Polypodium cyathisorum, Polypodium cyclocolpon,Polypodium daguense, Polypodium damunense, Polypodium dareiformioides,Polypodium dasypleura, Polypodium decipiens, Polypodium decorum,Polypodium delicatulum, Polypodium deltoideum, Polypodium demeraranum,Polypodium denticulatum, Polypodium diaphanum, Polypodium dilatatum,Polypodium dispersum, Polypodium dissectum, Polypodium dissimulans,Polypodium dolichosorum, Polypodium dolorense, Polypodiumdonnell-smithii, Polypodium drymoglossoides, Polypodium ebeninum,Polypodium eggersii, Polypodium elmeri, Polypodium elongatum, Polypodiumenterosoroides, Polypodium erubescens, Polypodium erythrolepis,Polypodium erythrotrichum, Polypodium eurybasis, Polypodium eurybasisvar. villosum, Polypodium exornans, Polypodium falcoideum, Polypodiumfallacissimum, Polypodium farinosum, Polypodium faucium, Polypodiumfeei, Polypodium ferrugineum, Polypodium feuillei, Polypodium firmulum,Polypodium firmum, Polypodium flaccidum, Polypodium flagellare,Polypodium flexuosum, Polypodium flexuosum var. ekmanii, Polypodiumforbesii, Polypodium formosanum, Polypodium fraxinifolium subsp.articulatum, Polypodium fraxinifolium subsp. luridum, Polypodiumfructuosum, Polypodium fucoides, Polypodium fulvescens, Polypodiumgaleottii, Polypodium glaucum, Polypodium glycyrrhiza, Polypodiumgracillimum, Polypodium gramineum, Polypodium grandifolium, Polypodiumgratum, Polypodium graveolens, Polypodium griseo-nigrum, Polypodiumgriseum, Polypodium guttatum, Polypodium haalilioanum, Polypodiumhammatisorum, Polypodium hancockii, Polypodium haplophlebicum,Polypodium harrisii, Polypodium hastatum var. simplex, Polypodiumhawaiiense, Polypodium heanophyllum, Polypodium helleri, Polypodiumhemionitidium, Polypodium henryi, Polypodium herzogii, Polypodiumhesperium, Polypodium hessii, Polypodium hombersleyi, Polypodiumhostmannii, Polypodium humile, Polypodium hyalinum, Polypodium iboense,Polypodium induens var. subdentatum, Polypodium insidiosum, Polypodiuminsigne, Polypodium intermedium subsp. masafueranum var. obtuseserratum,Polypodium intramarginale, Polypodium involutum, Polypodium itatiayense,Polypodium javanicum, Polypodium juglandifolium, Polypodium kaniense,Polypodium knowltoniorum, Polypodium kyimbilense, Polypodiuml'herminieri var. costaricense, Polypodium lachniferum f. incurvata,Polypodium lachniferum var. glabrescens, Polypodium lachnopus,Polypodium lanceolatum var. complanatum, Polypodium lanceolatum var.trichophorum, Polypodium latevagans, Polypodium laxifrons, Polypodiumlaxifrons var. lividum, Polypodium lehmannianum, Polypodium leiorhizum,Polypodium leptopodon, Polypodium leuconeuron var. angustifolia,Polypodium leuconeuron var. latifolium, Polypodium leucosticta,Polypodium limulum, Polypodium lindigii, Polypodium lineatum, Polypodiumlomarioides, Polypodium longifrons, Polypodium loretense, Polypodiumloriceum var. umbraticum, Polypodium loriforme, Polypodium loxogramme f.gigas, Polypodium ludens, Polypodium luzonicum, Polypodium lycopodioidesf. obtusum, Polypodium lycopodioides L., Polypodium macrolepis,Polypodium macrophyllum, Polypodium macrosorum, Polypodiummacrosphaerum, Polypodium maculosum, Polypodium madrense, Polypodiummanmeiense, Polypodium margaritiferum, Polypodium maritimum, Polypodiummartensii, Polypodium mayoris, Polypodium megalolepis, Polypodiummelanotrichum, Polypodium menisciifolium var. pubescens, Polypodiummeniscioides, Polypodium merrillii, Polypodium mettenii, Polypodiummexiae, Polypodium microsorum, Polypodium militare, Polypodium minimum,Polypodium minusculum, Polypodium mixtum, Polypodium mollendense,Polypodium mollissimum, Polypodium moniliforme var. minus, Polypodiummonoides, Polypodium monticola, Polypodium montigenum, Polypodiummoritzianum, Polypodium moultonii, Polypodium multicaudatum, Polypodiummultilineatum, Polypodium multisorum, Polypodium munchii, Polypodiummuscoides, Polypodium myriolepis, Polypodium myriophyllum, Polypodiummyriotrichum, Polypodium nematorhizon, Polypodium nemorale, Polypodiumnesioticum, Polypodium nigrescentium, Polypodium nigripes, Polypodiumnigrocinctum, Polypodium nimbatum, Polypodium nitidissimum, Polypodiumnitidissimum var. latior, Polypodium nubrigenum, Polypodium oligolepis,Polypodium oligosorum, Polypodium oligosorum, Polypodium olivaceum,Polypodium olivaceum var. elatum, Polypodium oodes, Polypodiumoosphaerum, Polypodium oreophilum, Polypodium ornatissimum, Polypodiumornatum, Polypodium ovatum, Polypodium oxylobum, Polypodium oxypholis,Polypodium pakkaense, Polypodium pallidum, Polypodium palmatopedatum,Polypodium palmeri, Polypodium panamense, Polypodium parvum, Polypodiumpatagonicum, Polypodium paucisorum, Polypodium pavonianum, Polypodiumpectinatum var. caliense, Polypodium pectinatum var. hispidum,Polypodium pellucidum, Polypodium pendulum var. boliviense, Polypodiumpercrassum, Polypodium perpusillum, Polypodium peruvianum var.subgibbosum, Polypodium phyllitidis var. elongatum, Polypodiumpichinchense, Polypodium pilosissimum, Polypodium pilosissimum var.glabriusculum, Polypodium pilossimum var. tunguraquensis, Polypodiumpityrolepis, Polypodium platyphyllum, Polypodium playfairii, Polypodiumplebeium var. cooperi, Polypodium plectolepidioides, Polypodiumpleolepis, Polypodium plesiosorum var.i, Polypodium podobasis,Polypodium podocarpum, Polypodium poloense, Polypodium polydatylon,Polypodium polypodioides var. aciculare, Polypodium polypodioides var.michauxianum, Polypodium praetermissum, Polypodium preslianum var.immersum, Polypodium procerum, Polypodium procerum, Polypodiumproductum, Polypodium productum, Polypodium prolongilobum, Polypodiumpropinguum, Polypodium proteus, Polypodium pruinatum, Polypodiumpseudocapillare, Polypodium pseudofratemum, Polypodium pseudonutans,Polypodium pseudoserratum, Polypodium pulcherrimum, Polypodiumpulogense, Polypodium pungens, Polypodium purpusii, Polypodium radicale,Polypodium randallii, Polypodium ratiborii, Polypodium reclinatum,Polypodium recreense, Polypodium repens var. abruptum, Polypodiumrevolvens, Polypodium rhachipterygium, Polypodium rhomboideum,Polypodium rigens, Polypodium robustum, Polypodium roraimense,Polypodium roraimense, Polypodium rosei, Polypodium rosenstockii,Polypodium rubidum, Polypodium rudimentum, Polypodium rusbyi, Polypodiumsablanianum, Polypodium sarmentosum, Polypodium saxicola, Polypodiumschenckii, Polypodium schlechteri, Polypodium scolopendria, Polypodiumscolopendria, Polypodium scolopendrium, Polypodium scouleri, Polypodiumscutulatum, Polypodium segregatum, Polypodium semihirsutum, Polypodiumsemihirsutum var. fuscosetosum, Polypodium senile var. minor, Polypodiumsericeolanatum, Polypodium serraeforme, Polypodium serricula, Polypodiumsesquipedala, Polypodium sessilifolium, Polypodium setosum var. calvum,Polypodium setulosum, Polypodium shaferi, Polypodium sibomense,Polypodium siccum, Polypodium simacense, Polypodium simulans, Polypodiumsingeri, Polypodium sinicum, Polypodium sintenisii, Polypodium skutchii,Polypodium sloanei, Polypodium sodiroi, Polypodium sordidulum,Polypodium sordidum, Polypodium sphaeropteroides, Polypodium sphenodes,Polypodium sprucei, Polypodium sprucei var. furcativenosa, Polypodiumsteirolepis, Polypodium stenobasis, Polypodium stenolepis, Polypodiumstenopterum, Polypodium subcapillare, Polypodium subflabelliforme,Polypodium subhemionitidium, Polypodium subinaequale, Polypodiumsubintegrum, Polypodium subspathulatum, Polypodium subtile, Polypodiumsubvestitum, Polypodium subviride, Polypodium superficiale var.attenuatum, Polypodium superficiale var. chinensis, Polypodiumsursumcurrens, Polypodium tablazianum, Polypodium taenifolium,Polypodium tamandarei, Polypodium tatei, Polypodium tenuiculum var.acrosora, Polypodium tenuiculum var. brasiliense, Polypodium tenuilore,Polypodium tenuinerve, Polypodium tepuiense, Polypodium teresae,Polypodium tetragonum var. incompletum, Polypodium thysanolepis var.bipinnatifidum, Polypodium thyssanolepis, var. thyssanolepis, Polypodiumthyssanolepsi, Polypodium tobagense, Polypodium trichophyllum,Polypodium tridactylum, Polypodium tridentatum, Polypodium trifurcatumvar. brevipes, Polypodium triglossum, Polypodium truncatulum, Polypodiumtruncicola var. major, Polypodium truncicola var. minor, Polypodiumtuberosum, Polypodium tunguraguae, Polypodium turquinum, Polypodiumturrialbae, Polypodium ursipes, Polypodium vagans, Polypodiumvaldealatum, Polypodium versteegii, Polypodium villagranii, Polypodiumvirginianum f. cambroideum, Polypodium virginianum f. peraferens,Polypodium vittarioides, Polypodium vulgare, Polypodium vulgare L.,Polypodium vulgare subsp. oreophilum, Polypodium vulgare var.acuminatum, Polypodium vulpinum, Polypodium williamsii, Polypodiumwobbense, Polypodium x fallacissimum-guttatum, Polypodium xantholepis,Polypodium xiphopteris, Polypodium yarumalense, Polypodium yungense, andPolypodium zosteriforme.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Nephrolepidaceae.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Nephrolepidaceae, GenusNephrolepis selected from, but not limited to, Nephrolepis abrupta,Nephrolepis acutifolia, Nephrolepis averyi, Nephrolepis biserrata,Nephrolepis brownii, Nephrolepis copelandi, Nephrolepis cordifolia,Nephrolepis davalliae, Nephrolepis davallioides, Nephrolepisdicksonioides, Nephrolepis exaltata, Nephrolepis falcata, Nephrolepisfalciformis, Nephrolepis hippocrepicis, Nephrolepis laurifolia,Nephrolepis lauterbachii, Nephrolepis medlerae, Nephrolepis obliterata,Nephrolepis pectinata, Nephrolepis pendula, Nephrolepis pseudobiserrata,Nephrolepis radicans, Nephrolepis rivularis, and Nephrolepis undulata.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the order Polypodiales, Family Polypodiaceae, Genus Colysisselected from, but not limited to, Colysis ampla, Colysis digitata,Colysis diversifolia, Colysis elegans Colysis elliptica, Colysisflexiloba, Colysis hemionitidea, Colysis hemitoma, Colysis henryi,Colysis insignis, Colysis intermedia, Colysis leveillei, Colysislongipes, Colysis pedunculata, Colysis pentaphylla, Colysis pothifolia,Colysis pteropus, Colysis shintenensis, Colysis simplicifrons, Colysistriphylla, Colysis wrightii, and Colysis x shintenensis.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Tectariaceae.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Tectariaceae, Genus Tectariaselected from, but not limited to, Tectaria acerifolia, Tectariaacrocarpa, Tectaria adenophora, Tectaria aequatoriensis, Tectariaamblyotis, Tectaria amphiblestra, Tectaria andersonii, Tectariaangelicifolia, Tectaria angulata, Tectaria antioquiana, Tectariaathyrioides, Tectaria athyriosora, Tectaria aurita, Tectaria balansae,Tectaria barberi, Tectaria barteri, Tectaria beccariana, Tectariablumeana, Tectaria brachiata, Tectaria brauniana, Tectaria brevilobata,Tectaria brooksii, Tectaria buchtienii, Tectaria calcarea, Tectariacamerooniana, Tectaria chattagramica, Tectaria cherasica, Tectariachimborazensis, Tectaria chinensis, Tectaria christii, Tectariachristovalensis, Tectaria cicutaria, Tectaria coadunata, Tectariaconfluens, Tectaria consimilis, Tectaria cordulata, Tectariacoriandrifolia, Tectaria craspedocarpa, Tectaria crenata, Tectariacrinigera, Tectaria croftii, Tectaria curtisii, Tectaria danfuensis,Tectaria decaryana, Tectaria decastroi, Tectaria decurrens, Tectariadegeneri, Tectaria dolichosora, Tectaria draconoptera, Tectaria dubia,Tectaria durvillei, Tectaria ebenina, Tectaria estremerana, Tectariaexauriculata, Tectaria fauriei, Tectaria fengii, Tectaria fernandensis,Tectaria ferruginea, Tectaria filisquamata, Tectaria fimbriata, Tectariafissa, Tectaria gaudichaudii, Tectaria gemmifera, Tectaria godeffroyi,Tectaria grandidentata, Tectaria griffithii var. singaporeana, Tectariagrossedentata, Tectaria hederifolia, Tectaria hekouensis, Tectariaheracleifolia, Tectaria herpetocaulos, Tectaria heterocarpa, Tectariahilocarpa, Tectaria holttumii, Tectaria hookeri, Tectaria humbertiana,Tectaria hymenodes, Tectaria hymenophylla, Tectaria impressa, Tectariaincisa, Tectaria inopinata, Tectaria isomorpha, Tectaria jacobsii,Tectaria jardini, Tectaria johannis-winkleri, Tectaria keckii, Tectariakehdingiana, Tectaria kingii, Tectaria kouniensis, Tectariakweichowensis, Tectaria labrusca, Tectaria lacei, Tectaria laotica,Tectaria latifolia, Tectaria lawrenceana, Tectaria laxa, Tectarialeptophylla, Tectaria lifuensis, Tectaria lizarzaburui, Tectaria lobbii,Tectaria lombokensis, Tectaria macrosora, Tectaria macrota, Tectariamadagascarica, Tectaria magnifica, Tectaria manilensis, Tectariamarchionica, Tectaria media, Tectaria melanocaulis, Tectariamelanocauloides, Tectaria melanorachis, Tectaria menyanthidis, Tectariamesodon, Tectaria mexicana, Tectaria microchlamys, Tectaria microlepis,Tectaria minuta, Tectaria moorei, Tectaria morlae, Tectaria moussetii,Tectaria murrayi, Tectaria nabirensis, Tectaria nausoriensis, Tectarianebulosa, Tectaria nesiotica, Tectaria nicaraguensis, Tectarianicotianifolia, Tectaria nitens, Tectaria novoguineensis, Tectariaorganensis, Tectaria palmate, Tectaria pandurifolia, Tectaria pedata,Tectaria pentagonalis, Tectaria perdimorpha, Tectaria phaeocaulis,Tectaria pica, Tectaria pilosa, Tectaria plantaginea, Tectariapleiosora, Tectaria pleiotoma, Tectaria poilanei, Tectaria polymorpha,Tectaria prolifera, Tectaria pseudosinuata, Tectariax pteropus-minor,Tectaria pubens, Tectaria puberula, Tectaria pubescens, Tectariaquinquefida, Tectaria quitensis, Tectaria ramosii, Tectaria rara,Tectaria remotipinna, Tectaria repanda, Tectaria rheophytica, Tectariarigida, Tectaria rivalis, Tectaria rockii, Tectaria rufescens, Tectariarufovillosa, Tectaria sagenioides, Tectaria schmutzii, Tectariaschultzei, Tectaria seemannii, Tectaria semibipinnata, Tectariasemipinnata, Tectaria seramensis, Tectaria siifolia, Tectariasimaoensis, Tectaria simonsii, Tectaria simulans, Tectaria singaporeana,Tectaria sinuata, Tectaria squamipes, Tectaria stalactica, Tectariastearnsii, Tectaria stenosemioides, Tectaria subcaudata, Tectariasubconfluens, Tectaria subcordata, Tectaria subdigitata, Tectariasubebenea, Tectaria subrepanda, Tectaria subsageniacea, Tectariasubtriloba, Tectaria subtriphylla, Tectaria sulitii, Tectaria suluensis,Tectaria sumatrana, Tectaria tabonensis, Tectaria taccifolia, Tectariatahitensis, Tectaria tenerifrons, Tectaria tenuifolia, Tectariateratocarpa, Tectaria ternata, Tectaria transiens, Tectaria translucens,Tectaria tricuspis, Tectaria trifida, Tectaria trifoliata, Tectariatriglossa, Tectaria triloba, Tectaria trimenii, Tectaria trinitensis,Tectaria tripartita, Tectaria variabilis, Tectaria vasta, Tectariavieillardii, Tectaria villosa, Tectaria vitiensis, Tectaria vivipara,Tectaria waterlotii, Tectaria weberi, Tectaria wightii, Tectariaxamesiana, Tectariax cynthiae, Tectaria yunnanensis, Tectaria zeylanica,and Tectaria zollingeri.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Davalliaceae.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Davalliaceae Genus Davalliaselected from, but not limited to, Davallia adiantoides, Davalliaamabilis, Davallia assamica, Davallia austrosinica, Davallia biflora,Davallia boryana, Davallia brachypoda, Davallia brevisora, Davalliabullata, Davallia bullata, Davallia calvescens, Davallia calvescens,Davallia canariensis, Davallia chaerophylla, Davallia chaerophylloide,Davallia chrysanthemifolia, Davallia clarkei, Davallia cumingii,Davallia cylindrica, Davallia divaricata, Davallia divaricata, Davalliadivaricata var. orientate, Davallia domingensis, Davallia dubia,Davallia elmeri, Davallia falcata, Davallia falcinella, Davalliaferulacea, Davallia flaccida, Davallia formosana, Davallia fumarioides,Davallia goudotiana, Davallia gracilis, Davallia griffithiana, Davalliagriffithiana, Davallia henryana, Davallia heterophylla, Davalliahookeriana, Davallia hymenophylloides, Davallia immersa, Davalliainaequalis var. minor, Davallia jamaicensis, Davallia khasiyana,Davallia kurzii, Davallia lepida, Davallia lepida, Davallia macraeana,Davallia magellanica, Davallia mariesii, Davallia membranulosa, Davalliamembranulosa, Davallia millefolium, Davallia moorei, Davalliamultidentata, Davallia nodosa, Davallia novae-guineae, Davalliaorientalis, Davallia parallela, Davallia parkeri, Davallia parvipinnula,Davallia patens, Davallia pectinata, Davallia perdurans, Davalliapilosula, Davallia platylepis, Davallia polypodioides, Davalliapolypodioides var. hispida, Davallia polypodioides var. pilosula,Davallia pseudocystopteris, Davallia puberula, Davallia pyramidata,Davallia pyxidata, Davallia repens, Davallia rhomboidea, Davalliarhomboidea, Davallia rhomboidea, Davallia sinensis, Davallia sloanei,Davallia solida, Davallia solida, Davallia stipellata, Davalliastrigosa, Davallia strigosa, Davallia strigosa var. rhomboidea, Davalliasubalpina, Davallia subsolida, Davallia teyermannii, Davalliatriangularis, Davallia tripinnata, Davallia truncata, Davalliatyermanni, Davallia tyermannii, Davallia uncinella, Davallia urophylla,Davallia vestita, Davallia wilfordii var. contracta, and Davalliayunnanensis.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Dryopteridaceae.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Dryopteridaceae, GenusPolystichum selected from, but not limited to, Polystichumacanthophyllum, Polystichum aculeatum, Polystichum acutidens,Polystichum acutipinnulum, Polystichum adungense, Polystichum alcicome,Polystichum altum, Polystichum anomalum, Polystichum ariticulatipilosum,Polystichum assurgentipinnum, Polystichum atkinsonii, Polystichumattenuatum, Polystichum auriculum, Polystichum bakerianum, Polystichumbaoxingense, Polystichum biaristatum, Polystichum bifidum, Polystichumbigemmatum, Polystichum bissectum, Polystichum bomiense, Polystichumbrachypterum, Polystichum braunii, Polystichum capillipes, Polystichumcastaneum, Polystichum chingiae, Polystichum christii, Polystichumchunii, Polystichum consimile, Polystichum costularisorum, Polystichumcraspedosorum, Polystichum crassinervium, Polystichum cringerum,Polystichum cuneatiforme, Polystichum cyclolobum, Polystichumdaguanense, Polystichum dangii, Polystichum delavayi, Polystichumdeltodon, Polystichum dielsii, Polystichum diffundens, Polystichumdiscretum, Polystichum disjunctum, Polystichum duthiei, Polystichumelevatovenusum, Polystichum erosum, Polystichum exauriforme, Polystichumexcellens, Polystichum excelsius, Polystichum fimbriatum, Polystichumformosanum, Polystichum frigidicola, Polystichum fugongense, Polystichumgongboense, Polystichum grandifrons, Polystichum guangxiense,Polystichum gymnocarpium, Polystichum habaense, Polystichum hancockii,Polystichum hecatopteron, Polystichum herbaceum, Polystichumhouchangense, Polystichum huae, Polystichum ichangense, Polystichuminaense, Polystichum incisopinnulum, Polystichum integrilimbum,Polystichum integrilobum, Polystichum jinfoshaense, Polystichumjiulaodongense, Polystichum jizhushanense, Polystichum kangdingense,Polystichum kungianum, Polystichum kwangtungense, Polystichumlachenense, Polystichum lanceolatum, Polystichum langchungense,Polystichum latilepis, Polystichum lentum, Polystichum leveillei,Polystichum liui, Polystichum lonchitis, Polystichum longiaristatum,Polystichum longidens, Polystichum longipaleatum, Polystichum longipes,Polystichum longipinnulum, Polystichum longispinosum, Polystichumlongissimum, Polystichum macrochlaenum, Polystichum makinoi, Polystichummanmeiense, Polystichum martinii, Polystichum mayebarae, Polystichummedogense, Polystichum mehrae, Polystichum meiguense, Polystichummelanostipes, Polystichum mollissimum, Polystichum morii, Polystichummoupinense, Polystichum muscicola, Polystichum nayongense, Polystichumneoliuii, Polystichum neolobatum, Polystichum nepalense, Polystichumnigrum, Polystichum ningshenense, Polystichum nudisorum, Polystichumobliquum, Polystichum oblongum, Polystichum oligocarpum, Polystichumomeiense, Polystichum oreodoxa, Polystichum orientalitibeticum,Polystichum otophorum, Polystichum ovato-paleaceum, Polystichumparamoupinense, Polystichum parvifoliolatum, Polystichum parvipinnulum,Polystichum pianmaense, Polystichum piceo-paleaceum, Polystichumpolyblepharum, Polystichum prescottianum, Polystichum prionolepis,Polystichum pseudocastaneum, Polystichum pseudolanceolatum, Polystichumpseudomakinoi, Polystichum pseudorhomboideum, Polystichum pseudosetosum,Polystichum pseudoxiphophyllum, Polystichum punctiferum, Polystichumputeicola, Polystichum pycnopterum, Polystichum qamdoense, Polystichumretrosopaleaceum, Polystichum revolutum, Polystichum rhombiforme,Polystichum rigens, Polystichum robustum, Polystichum rufopaleaceum,Polystichum saxicola, Polystichum semifertile, Polystichum setillosum,Polystichum shandongense, Polystichum shensiense, Polystichum shimurae,Polystichum simplicipinnum, Polystichum sinense, Polystichumsinotsus-simense, Polystichum sozanense, Polystichum speluncicola,Polystichum squarrosum, Polystichum stenophyllum, Polystichum stimulans,Polystichum subacutidens, Polystichum subdeltodon, Polystichumsubfimbriatum, Polystichum submarginale, Polystichum submite,Polystichum subulatum, Polystichum tacticopterum, Polystichumtaizhongense, Polystichum tangmaiense, Polystichum thomsonii,Polystichum tibeticum, Polystichum tonkinense, Polystichum tripteron,Polystichum tsingkanshanense, Polystichum tsus-simense, Polystichumwattii, Polystichum xiphophyllum, Polystichum yadongense, Polystichumyuanum, Polystichum yunnanense, and Polystichum zayuense.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Pteridaceae, Genus Adiantaceaeselected from, but not limited to, Adiantum aethiopicum, Adiantumaleuticum, Adiantum bonatianum, Adiantum cajennense, Adiantumcapillus-junonis, Adiantum capillus-veneris, Adiantum caudatum, Adiantumchienii, Adiantum chilense, Adiantum cuneatum, Adiantum cunninghamii,Adiantum davidii, Adiantum diaphanum, Adiantum edentulum, Adiantumedgeworthii, Adiantum excisum, Adiantum fengianum, Adiantum fimbriatum,Adiantum flabellulatum, Adiantum formosanum, Adiantum formosum, Adiantumfulvum, Adiantum gravesii, Adiantum hispidulum, Adiantum induratum,Adiantum jordanii, Adiantum juxtapositum, Adiantum latifolium, Adiantumleveillei, Adiantum lianxianense, Adiantum malesianum, Adiantummariesii, Adiantum monochlamys, Adiantum myriosorum, Adiantum obliquum,Adiantum ogasawarense, Adiantum pedatum, Adiantum pentadactylon,Adiantum peruvianum, Adiantum philippense, Adiantum princeps, Adiantumpubescens, Adiantum raddianum, Adiantum reniforme, Adiantum roborowskii,Adiantum serratodentatum, Adiantum sinicum, Adiantum soboliferum,Adiantum subcordatum, Adiantum tenerum, Adiantum terminatum, Adiantumtetraphyllum, Adiantum venustum, Adiantum viridescens, and Adiantumviridimontanum. In some embodiments, the IPD113 polypeptide is derivedfrom a fern species in the

Order Polypodiales, Family Aspleniaceae, Genus Asplenium. In someembodiments, the nucleic acid molecule encoding the IPD113 polypeptideis derived from a fern species in the Order Polypodiales, FamilyAspleniaceae, Genus Asplenium L selected from, but not limited to,Asplenium abbreviatum, Asplenium abrotanoides, Asplenium abscissum var.subaequilaterale, Asplenium abscissum, Asplenium achilleifolium,Asplenium acuminatum, Asplenium adiantifrons, Asplenium adiantoides,Asplenium adiantoides var. squamulosum, Asplenium adiantum-nigrum L.,Asplenium adiantum-nigrum var. adiantum-nigrum, Aspleniumadiantum-nigrum var. yuanum, Asplenium adnatum, Asplenium aethiopicum,Asplenium affine, Asplenium affine var. affine, Asplenium affine var.gilpinae, Asplenium affine var. mettenii, Asplenium affine var. pecten,Asplenium africanum, Asplenium afzelii, Asplenium aitchisonii, Aspleniumalatulum, Asplenium alatum, Asplenium alfredii, Asplenium altajense,Asplenium amabile, Asplenium ambohitantelense, Asplenium anceps var.proliferum, Asplenium andapense, Asplenium andersonii, Aspleniumangustatum, Asplenium angustum, Asplenium anisophyllum, Aspleniumannetii, Asplenium antiquum, Asplenium antrophyoides, Asplenium apertum,Asplenium apogamum, Asplenium aquaticum, Asplenium arboreum, Aspleniumarcanum, Asplenium arcuatum, Asplenium argentinum, Asplenium argutum,Asplenium aspidiiforme, Asplenium aspidioides, Asplenium asterolepis,Asplenium auricularium var. acutidens, Asplenium auricularium var.subintegerrimum, Asplenium auriculatum, Asplenium auriculatum var.aequilaterale, Asplenium auritum, Asplenium auritum var. auriculatum,Asplenium auritum var. auritum, Asplenium auritum var. bipinnatifidum,Asplenium auritum var. bipinnatisectum, Asplenium auritum var.davallioides, Asplenium auritum var. macilentum, Asplenium auritum var.rigidum, Asplenium auritum var. subsimplex, Asplenium austrochinense,Asplenium ayopayense, Asplenium badinii, Asplenium balense, Aspleniumballivianii, Asplenium bangii, Asplenium bangii, Asplenium barbaense,Asplenium barclayanum, Asplenium barkamense, Asplenium barteri,Asplenium basiscopicum, Asplenium bicrenatum, Asplenium bifrons,Asplenium bipartitum, Asplenium blastophorum, Asplenium blepharodes,Asplenium blepharophorum, Asplenium boiteaui, Asplenium bolivianum,Asplenium boltonii, Asplenium borealichinense, Asplenium bradei,Asplenium bradeorum, Asplenium bradleyi, Asplenium brausei, Aspleniumbreedlovei, Asplenium buettneri, Asplenium buettneri var. hildebrandtii,Asplenium bulbiferum, Asplenium bullatum var. bullatum, Aspleniumbullatum var. shikokianum, Asplenium bullatum, Asplenium cancellatum,Asplenium capillipes, Asplenium cardiophyllum (Hance), Aspleniumcaripense, Asplenium carvalhoanum, Asplenium castaneoviride, Aspleniumcastaneum, Asplenium caudatum, Asplenium celtidifolium (Kunze),Asplenium ceratolepis, Asplenium changputungense, Asplenium chaseanum,Asplenium cheilosorum, Asplenium chengkouense, Asplenium chihuahuense,Asplenium chimantae, Asplenium chimborazense, Asplenium chingianum,Asplenium chlorophyllum, Asplenium chondrophyllum, Asplenium cicutarium,Asplenium cicutarium var. paleaceum, Asplenium cirrhatum, Aspleniumcladolepton, Asplenium claussenii, Asplenium coenobiale, Aspleniumcommutatum, Asplenium congestum, Asplenium conquisitum, Aspleniumconsimile, Asplenium contiguum, Asplenium contiguum var. hirtulum,Asplenium corderoi, Asplenium cordovense, Asplenium coriaceum, Aspleniumcoriifolium, Asplenium correardii, Asplenium costale, Asplenium costalevar. robustum, Asplenium cowanii, Asplenium crenulatoserrulatum,Asplenium crenulatum, Asplenium crinicaule, Asplenium crinulosum,Asplenium cristatum, Asplenium cryptolepis Fernald, Aspleniumcultrifolium L., Asplenium cuneatiforme, Asplenium cuneatum, Aspleniumcurvatum, Asplenium cuspidatum, Asplenium cuspidatum var cuspidatum,Asplenium cuspidatum var. foeniculaceum, Asplenium cuspidatum var.triculum, Asplenium cuspidatum var. tripinnatum, Asplenium dalhousiae,Asplenium dareoides, Asplenium davallioides, Asplenium davisii,Asplenium debile, Asplenium debile, Asplenium decussatum, Aspleniumdelavayi, Asplenium delicatulum, Asplenium delicatulum var. cocosensis,Asplenium delitescens, Asplenium delitescens X laetum, Asplenium densum,Asplenium dentatum L., Asplenium dentatum L., Asplenium depauperatum,Asplenium deqenense, Asplenium dianae, Asplenium difforme, Aspleniumdilatatum, Asplenium dimidiatum, Asplenium dimidiatum var. boliviense,Asplenium diplazisorum, Asplenium dissectum, Asplenium distans,Asplenium divaricatum, Asplenium divergens, Asplenium divisissimum,Asplenium doederleinii, Asplenium donnell-smithii, Asplenium dregeanum,Asplenium dulongjiangense, Asplenium duplicatoserratum, Aspleniumeatonii, Asplenium ebeneum, Asplenium ebenoides, Asplenium ecuadorense,Asplenium eggersii, Asplenium emarginatum, Asplenium enatum, Aspleniumensiforme fo. bicuspe, Asplenium ensiforme fo. ensiforme, Aspleniumensiforme fo. stenophyllum, Asplenium ensiforme, Asplenium erectum var.erectum, Asplenium erectum var. gracile, Asplenium erectum var.usambarense, Asplenium erectum var. zeyheri, &, Asplenium erosum L.,Asplenium escaleroense, Asplenium esculentum, Asplenium eutecnum,Asplenium excelsum, Asplenium excisum, Asplenium exiguum, Aspleniumextensum, Asplenium falcatum, Asplenium falcinellum, Asplenium faurei,Asplenium feel, Asplenium fengyangshanense, Asplenium ferulaceum,Asplenium fibrillosum, Asplenium filix-femina, Asplenium finckii,Asplenium finlaysonianum, Asplenium flabellulatum, Aspleniumflabellulatum var flabellulatum, Asplenium flabellulatum var. partitum,Asplenium flaccidum, Asplenium flavescens, Asplenium flavidum, Aspleniumflexuosum, Asplenium fluminense, Asplenium foeniculaceum, Aspleniumformosanum, Asplenium formosum var. carolinum, Asplenium formosum var.incultum, Asplenium formosum, Asplenium foumieri, Asplenium fragile,Asplenium fragile var. lomense, Asplenium fragrans, Asplenium fragransvar. foeniculaceum, Asplenium franconis var. gracile, Aspleniumfraxinifolium, Asplenium friesiorum, Asplenium friesiorum var.nesophilum, Asplenium fugax, Asplenium fujianense, Asplenium furcatum,Asplenium furfuraceum, Asplenium fuscipes, Asplenium fuscopubescens,Asplenium galeottii, Asplenium gautieri, Asplenium gemmiferum, Aspleniumgentryi, Asplenium geppii, Asplenium ghiesbreghtii, Asplenium gilliesii,Asplenium gilpinae, Asplenium glanduliserratum, Asplenium glenniei,Asplenium goldmannii, Asplenium gomezianum, Asplenium grande, Aspleniumgrandifolium, Asplenium grandifrons, Asplenium gregoriae, Aspleniumgriffithianum, Asplenium gulingense, Asplenium hainanense, Aspleniumhallbergii, Asplenium hallei, Asplenium hallii, Asplenium hangzhouense,Asplenium haplophyllum, Asplenium harpeodes, Asplenium harpeodes var.glaucovirens, Asplenium harpeodes var. incisum, Asplenium harrisiiJenman, Asplenium harrisonii, Asplenium hastatum, Asplenium hebeiense,Asplenium hemionitideum, Asplenium hemitomum, Asplenium henryi,Asplenium herpetopteris, Asplenium herpetopteris var herpetopteris,Asplenium herpetopteris var. acutipinnata, Asplenium herpetopteris var.masoulae, Asplenium herpetopteris var. villosum, Asplenium hesperium,Asplenium heterochroum, Asplenium hians, Asplenium hians var.pallescens, Asplenium hoffmannii, Asplenium holophlebium, Aspleniumhondoense, Asplenium horridum, Asplenium hostmannii, Aspleniumhumistratum, Asplenium hypomelas, Asplenium inaequilaterale, Aspleniumincisum, Asplenium incurvatum, Asplenium indicum, Asplenium indicum var.indicum, Asplenium indicum var. yoshingagae, Asplenium induratum,Asplenium indusiatum, Asplenium inexpectatum, Asplenium insigne,Asplenium insiticium, Asplenium insolitum, Asplenium integerrimum,Asplenium interjectum, Asplenium jamesonii, Asplenium jaundeense,Asplenium juglandifolium, Asplenium kangdingense, Asplenium kansuense,Asplenium kassneri, Asplenium kaulfussii, Asplenium kellermanii,Asplenium kentuckiense, Asplenium khullarii, Asplenium kiangsuense,Asplenium kunzeanum, Asplenium lacerum, Asplenium laciniatum, Aspleniumlaciniatum var. acutipinna, Asplenium laciniatum var. laciniatum,Asplenium laetum fo. minor, Asplenium laetum, Asplenium laetum var.incisoserratum, Asplenium lamprocaulon, Asplenium laserpitiifolium var.morrisonense, Asplenium lastii, Asplenium latedens, Aspleniumlatifolium, Asplenium laui, Asplenium laurentii, Asplenium leandrianum,Asplenium lechleri, Asplenium leiboense, Asplenium lepidorachis,Asplenium leptochlamys, Asplenium leptophyllum, Asplenium levyi,Asplenium lindbergii, Asplenium lindeni, Asplenium lineatum, Aspleniumlividum, Asplenium lobatum, Asplenium lobulatum, Asplenium lokohoense,Asplenium longicauda, Asplenium longicaudatum, Asplenium longifolium,Asplenium longisorum, Asplenium longjinense, Asplenium lorentzii,Asplenium loriceum, Asplenium loxogrammoides, Asplenium lugubre,Asplenium lunulatum, Asplenium lunulatum var. pteropus, Aspleniumlushanense, Asplenium lydgatei, Asplenium macilentum, Asplenium macraei,Asplenium macrodictyon, Asplenium macrophlebium, Asplenium macrophyllum,Asplenium macropterum, Asplenium macrosorum, Asplenium macrotis,Asplenium macrurum, Asplenium mainlingense, Asplenium mangindranense,Asplenium mannii, Asplenium marginatum L., Asplenium marojejyense,Asplenium martianum, Asplenium matsumurae, Asplenium mauritiensisLorence, Asplenium maximum, Asplenium, ii, Asplenium megalura, Aspleniummegaphyllum, Asplenium meiotomum, Asplenium melanopus, Aspleniummembranifolium, Asplenium meniscioides, Asplenium mesosorum, Aspleniummexicanum, Asplenium micropaleatum, Asplenium microtum, Aspleniummildbraedii, Asplenium mildei, Asplenium minimum, Asplenium minutum,Asplenium miradorense, Asplenium miyunense, Asplenium moccenianum,Asplenium mocquerysii, Asplenium modestum, Asplenium monanthemum var.menziesii, Asplenium monanthes L., Asplenium monanthes var monanthes,Asplenium monanthes var. castaneum, Asplenium monanthes var. wagneri,Asplenium monanthes var. yungense, Asplenium monodon, Aspleniummontanum, Asplenium mosetenense, Asplenium moupinense, Aspleniummucronatum, Asplenium munchii, Asplenium muticum, Asplenium myapteron,Asplenium myriophyllu, Asplenium nakanoanum, Asplenium nanchuanense,Asplenium nemorale, Asplenium neolaserpitiifolium, Aspleniumneomutijugum, Asplenium neovarians, Asplenium nesii, Aspleniumnesioticum, Asplenium nidus L., Asplenium nigricans, Aspleniumniponicum, Asplenium normale, Asplenium normale var. angustum, Aspleniumobesum, Asplenium oblongatum, Asplenium oblongifolium, Aspleniumobovatum, Asplenium obscurum, Asplenium obscurum var. angustum,Asplenium obtusatum var. obtusatum, Asplenium obtusatum var. sphenoides,Asplenium obtusifolium L., Asplenium obtusissimum, Asplenium obversum,Asplenium ochraceum, Asplenium oellgaardii, Asplenium ofeliae, Aspleniumoldhami, Asplenium oligosorum, Asplenium olivaceum, Asplenium onopterisL., Asplenium onustum, Asplenium ortegae, Asplenium otites, Aspleniumpalaciosii, Asplenium palmeri, Asplenium partitum, Asplenium parvisorum,Asplenium parviusculum, Asplenium parvulum, Asplenium patens, Aspleniumpaucifolium, Asplenium paucijugum, Asplenium paucivenosum, Aspleniumpearcei, Asplenium pekinense, Asplenium pellucidum, Asplenium pendulum,Asplenium petiolulatum, Asplenium phyllitidis, Aspleniumpimpinellifolium, Asplenium pinnatifidum, Asplenium pinnatum, Aspleniumplatyneuron, Asplenium platyneuron var. bacculum-rubrum, Aspleniumplatyneuron var. incisum, Asplenium platyphyllum, Asplenium plumbeum,Asplenium poloense, Asplenium polymeris, Asplenium polymorphum,Asplenium polyodon, Asplenium polyodon var. knudsenii, Aspleniumpolyodon var. nitidulum, Asplenium polyodon var. sectum, Aspleniumpolyodon var. subcaudatum, Asplenium polyphyllum, Asplenium poolii,Asplenium poolii fo. simplex, Asplenium poolii var. linearipinnatum,Asplenium potosinum, Asplenium potosinum var. incisum, Aspleniumpraegracile, Asplenium praemorsum, Asplenium preussii, Aspleniumpringleanum, Asplenium pringlei, Asplenium prionitis, Aspleniumprocerum, Asplenium progrediens, Asplenium projectum, Aspleniumprolongatum, Asplenium propinquum, Asplenium protensum, Aspleniumpseudoangustum, Asplenium pseudoerectum, Asplenium pseudofontanum,Asplenium pseudolaserpitiifolium, Asplenium pseudonormale, Aspleniumpseudopellucidum, Asplenium pseudopraemorsum, Asplenium pseudovarians,Asplenium pseudowilfordii, Asplenium pseudowrightii, Aspleniumpsilacrum, Asplenium pteropus, Asplenium pubirhizoma, Aspleniumpulchellum, Asplenium pulchellum var. subhorizontale, Aspleniumpulcherrimum, Asplenium pulicosum, Asplenium pulicosum var. majus,Asplenium pululahuae, Asplenium pumilum, Asplenium pumilum var.hymenophylloides, Asplenium pumilum var. laciniatum, Aspleniumpurdieanum, Asplenium purpurascens, Asplenium pyramidatum, Aspleniumqiujiangense, Asplenium quercicola, Asplenium quitense, Aspleniumraddianum, Asplenium radiatum, Asplenium radicans L., Aspleniumradicans, Asplenium radicans var. costaricense, Asplenium radicans var.partitum, Asplenium radicans var. radicans, Asplenium radicans var.uniseriale, Asplenium recumbens, Asplenium reflexum, Asplenium regularevar. latior, Asplenium repandulum, Asplenium repens, Asplenium repente,Asplenium resiliens, Asplenium retusulum, Asplenium rhipidoneuron,Asplenium rhizophorum L., Asplenium rhizophyllum, Asplenium rhizophyllumL., Asplenium rhizophyllum var. proliferum, Asplenium rhomboideum,Asplenium rigidum, Asplenium riparium, Asplenium rivale, Aspleniumrockii, Asplenium roemerianum, Asplenium roemerianum var. mindensis,Asplenium rosenstockianum, Asplenium rubinum, Asplenium ruizianum,Asplenium rusbyanum, Asplenium ruta-muraria L., Asplenium ruta-murariavar. cryptolepis, Asplenium rutaceum, Asplenium rutaceum var.disculiferum, Asplenium rutaefolium, Asplenium rutifolium, Aspleniumsalicifolium L., Asplenium salicifolium var. aequilaterale, Aspleniumsalicifolium var. salicifolium, Asplenium sampsoni, Asplenium sanchezii,Asplenium sanderi, Asplenium sandersonii, Asplenium sanguinolentum,Asplenium sarelii, Asplenium sarelii var. magnum, Asplenium sarelii var.sarelii, Asplenium saxicola, Asplenium scalifolium, Aspleniumscandicinum, Asplenium schizophyllum, Asplenium schkuhrii, Aspleniumsciadophilum, Asplenium scolopendrium L., Asplenium scortechinii,Asplenium seileri, Asplenium semipinnatum, Asplenium septentrionale,Asplenium serra, Asplenium serra var. imrayanum, Aspleniumserratissimum, Asplenium serratum L., Asplenium serratum var. caudatum,Asplenium serricula, Asplenium sessilifolium, Asplenium sessilifoliumvar. guatemalense, Asplenium sessilifolium var. minus, Aspleniumsessilifolium var. occidentale, Asplenium sessilipinnum, Aspleniumsetosum, Asplenium shepherdii, Asplenium shepherdii var. bipinnatum,Asplenium shepherdii var. flagelliferum, Asplenium shikokianum,Asplenium simii, Asplenium simonsianum, Asplenium sintenisii, Aspleniumskinneri, Asplenium skinneri, Asplenium sodiroi, Aspleniumsoleirolioides, Asplenium solidum var. stenophyllum, Asplenium solmsii,Asplenium sp.-N.-Halle-2234, Asplenium spathulinum, Aspleniumspectabile, Asplenium speluncae, Asplenium sphaerosporum, Aspleniumsphenotomum, Asplenium spinescens, Asplenium splendens, Aspleniumsprucei, Asplenium squamosum L., Asplenium standleyi, Aspleniumstellatum, Asplenium stenocarpum, Asplenium stoloniferum, Aspleniumstolonipes, Asplenium striatum L., Asplenium stuebelianum, Aspleniumstuhlmannii, Asplenium suave, Asplenium subalatum, Aspleniumsubcrenatum, Asplenium subdigitatum, Asplenium subdimidiatum, Aspleniumsubintegrum, Asplenium sublaserpitiifolium, Asplenium sublongum,Asplenium subnudum, Asplenium suborbiculare, Asplenium subtenuifolium,Asplenium subtile, Asplenium subtoramanum, Asplenium subtrapezoideum,Asplenium subvarians, Asplenium sulcatum, Asplenium sylvaticum,Asplenium szechuanense, Asplenium taiwanense, Asplenium tenerrimum,Asplenium tenerum, Asplenium tenuicaule, Asplenium tenuifolium,Asplenium tenuifolium var. minor, Asplenium tenuifolium var.tenuifolium, Asplenium tenuissimum, Asplenium ternatum, Aspleniumtheciferum, Asplenium theciferum var. concinnum, Asplenium thunbergii,Asplenium tianmushanense, Asplenium tianshanense, Asplenium tibeticum,Asplenium tocoraniense, Asplenium toramanum, Asplenium trapezoideum,Asplenium tricholepis, Asplenium trichomanes L., Asplenium trichomanessubsp. inexpectans, Asplenium trichomanes subsp. quadrivalens, Aspleniumtrichomanes subsp. trichomanes, Asplenium trichomanes var. harovii,Asplenium trichomanes var. herbaceum, Asplenium trichomanes var. repens,Asplenium trichomanes var. viridissimum, Asplenium trichomanes-dentatumL., Asplenium trigonopterum, Asplenium trilobatum, Asplenium trilobum,Asplenium triphyllum, Asplenium triphyllum var. compactum, Aspleniumtriphyllum var. gracillimum, Asplenium triphyllum var. herbaceum,Asplenium tripteropus, Asplenium triquetrum, Asplenium truncorum,Asplenium tsaratananense, Asplenium tucumanense, Asplenium tuerckheimii,Asplenium tunquiniense, Asplenium ulbrichtii, Asplenium ultimum,Asplenium unilaterale, Asplenium unilaterale var. decurrens, Aspleniumunilaterale var. udum, Asplenium unilaterale var. unilaterale, Aspleniumuniseriale, Asplenium uropteron, Asplenium vagans, Aspleniumvareschianum, Asplenium variabile var. paucijugum, Asplenium variabilevar. variabile, Asplenium varians subsp. fimbriatum, Asplenium varians,Asplenium vastum, Asplenium venturae, Asplenium venulosum, Aspleniumverapax, Asplenium vesiculosum, Asplenium vespertinum, Aspleniumvillosum, Asplenium virens, Asplenium viride, Asplenium viridifrons,Asplenium virillae, Asplenium viviparioides, Asplenium viviparum,Asplenium viviparum var viviparum, Asplenium viviparum var. lineatu,Asplenium volubile, Asplenium vulcanicum, Asplenium wacketii, Aspleniumwagneri, Asplenium wallichianum, Asplenium wameckei, Aspleniumwilfordii, Asplenium williamsii, Asplenium wrightii, Aspleniumwrightioides, Asplenium wuliangshanense, Asplenium xianqianense,Asplenium xinjiangense, Asplenium xinyiense, Asplenium yelagagense,Asplenium yoshinagae, Asplenium yunnanense, Asplenium zamiifolium,Asplenium zanzibaricum, Asplenium biscayneanum, Asplenium curtissii,Asplenium ebenoides, Asplenium herb-wagneri, Asplenium heteroresiliens,Asplenium kenzoi, Asplenium plenum, Asplenium wangii, and Aspleniumxclermontiae, Asplenium x gravesii.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.In some embodiments, the nucleic acid molecule encoding the IPD113polypeptide is derived from a fern species in the Order Polypodiales,Family Blechnaceae, Genus Blechnum L. selected from, but not limited to,Blechnum amabile, Blechnum appendiculatum, Blechnum articulatum,Blechnum australe, Blechnum austrobrasilianum, Blechnum binervatum,Blechnum blechnoides, Blechnum brasiliense, Blechnum capense, Blechnumcartilagineum, Blechnum castaneum, Blechnum chambersii, Blechnumchilense, Blechnum colensoi, Blechnum contiguum, Blechnum cordatum,Blechnum coriaceum, Blechnum discolor, Blechnum doodioides, Blechnumdurum, Blechnum eburneum, Blechnum ensiforme, Blechnum filiforme,Blechnum fluviatile, Blechnum fragile, Blechnum fraseri, Blechnumfullagari, Blechnum gibbum, Blechnum glandulosum, Blechnum gracile,Blechnum hancockii, Blechnum hastatum, Blechnum howeanum, Blechnumindicum, Blechnum kunthianum, Blechnum laevigatum, Blechnum loxense,Blechnum magellanicum, Blechnum membranaceum, Blechnum microbasis,Blechnum microphyllum, Blechnum milnei, Blechnum minus, Blechnummochaenum, Blechnum montanum, Blechnum moorei, Blechnum moritzianum,Blechnum nigrum, Blechnum niponicum, Blechnum norfolkianum, Blechnumnovae-zelandiae, Blechnum nudum, Blechnum obtusatum, Blechnumoccidentale, Blechnum oceanicum, Blechnum orientale, Blechnumpatersonii, Blechnum penna-marina, Blechnum polypodioides, Blechnumprocerum, Blechnum punctulatum, Blechnum sampaioanum, Blechnumschiedeanum, Blechnum schomburgkii, Blechnum serrulatum, Blechnumsimillimum, Blechnum spicant, Blechnum stipitellatum, Blechnum tabulare,Blechnum triangularifolium, Blechnum vieillardii, Blechnum vulcanicum,Blechnum wattsii, Blechnum whelanii, and Blechnum wurunuran.

In some embodiments, the nucleic acid encoding the IPD113 polypeptide isderived from a fern species in the Order Schizaeales; FamilySchizaeaceae, Genus Lygodium selected from, but not limited to, Lygodiumarticulatum, Lygodium circinatum, Lygodium conforme, Lygodium cubense,Lygodium digitatum, Lygodium flexuosum, Lygodium heterodoxum, Lygodiumjaponicum, Lygodium kerstenii, Lygodium lanceolatum, Lygodiumlongifolium, Lygodium merrilii, Lygodium micans, Lygodium microphyllum,Lygodium microstachyum, Lygodium oligostachyum, Lygodium palmatum,Lygodium polystachyum, Lygodium radiatum, Lygodium reticulatum, Lygodiumsalicifolium, Lygodium scandens, Lygodium smithianum, Lygodiumsubareolatum, Lygodium trifurcatum, Lygodium venustum, Lygodiumversteeghii, Lygodium volubile, and Lygodium yunnanense.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Genus Ophioglossum L., Botrychium, Botrypus,Helminthostachys, Ophioderma, Cheiroglossa, Sceptridium or Mankyua. Insome embodiments, the IPD113 polypeptide is derived from a fern speciesin the Ophioglossum Genus is selected from, but not limited to,Ophioglossum californicum, Ophioglossum coriaceum, Ophioglossumcostatum, Ophioglossum crotalophoroides, Ophioglossum engelmannii,Ophioglossum falcatum, Ophioglossum gomezianum, Ophioglossum gramineum,Ophioglossum kawamurae, Ophioglossum lusitanicum, Ophioglossumnamegatae, Ophioglossum nudicaule, Ophioglossum palmatum, Ophioglossumparvum, Ophioglossum pedunculosum, Ophioglossum pendulum, Ophioglossumpetiolatum, Ophioglossum pusillum, Ophioglossum reticulatum,Ophioglossum richardsiae, Ophioglossum thermale, and Ophioglossumvulgatum.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Pyrrosia Genus selected from, but not limited to,Pyrrosia abbreviata, Pyrrosia angustata, Pyrrosia angustissima, Pyrrosiaassimilis, Pyrrosia asterosora, Pyrrosia blepharolepis, Pyrrosiaboothii, Pyrrosia borneensis, Pyrrosia brassii, Pyrrosia christii,Pyrrosia confluens, Pyrrosia costata, Pyrrosia dimorpha, Pyrrosiadispar, Pyrrosia distichocarpa, Pyrrosia drakeana, Pyrrosiaeleagnifolia, Pyrrosia fengiana, Pyrrosia flocculosa, Pyrrosiafoveolata, Pyrrosia fuohaiensis, Pyrrosia gardneri, Pyrrosia hastata,Pyrrosia heterophylla, Pyrrosia intermedia, Pyrrosia laevis, Pyrrosialanceolata, Pyrrosia liebuschii, Pyrrosia linearifolia, Pyrrosia lingua,Pyrrosia longifolia, Pyrrosia macrocarpa, Pyrrosia madagascariensis,Pyrrosia mannii, Pyrrosia matsudai, Pyrrosia mechowii, Pyrrosiamicraster, Pyrrosia morns, Pyrrosia novo-guineae, Pyrrosianummulariifolia, Pyrrosia oblanceolata, Pyrrosia obovata, Pyrrosiapannosa, Pyrrosia petiolosa, Pyrrosia piloselloides, Pyrrosiapolydactyla, Pyrrosia porosa, Pyrrosia princeps, Pyrrosiapseudodrakeana, Pyrrosia rasamalae, Pyrrosia rhodesiana, Pyrrosiarupestris, Pyrrosia samarensis, Pyrrosia scolopendrina, Pyrrosiasheareri, Pyrrosia shennongensis, Pyrrosia similis, Pyrrosiasphaerosticha, Pyrrosia stigmosa, Pyrrosia stolzii, Pyrrosiasubfurfuracea, Pyrrosia transmorrisonensis, and Pyrrosia tricholepis.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Doryopteris Genus selected from, but not limited to,Doryopteris collina, Doryopteris concolor, Doryopteris conformis,Doryopteris cordata, Doryopteris cordifolia, Doryopteris crenulans,Doryopteris cyclophylla, Doryopteris davidsei, Doryopteris decipiens,Doryopteris decora, Doryopteris effusa, Doryopteris humbertii,Doryopteris kirkii, Doryopteris kitchingii, Doryopteris latiloba,Doryopteris lomariacea, Doryopteris lorentzii, Doryopteris ludens,Doryopteris madagascariensis, Doryopteris michelii, Doryopteris nobilis,Doryopteris ornithopus, Doryopteris patens, Doryopteris patula,Doryopteris patula, Doryopteris pedata, Doryopteris pedata, Doryopterispedatoides, Doryopteris pilosa, Doryopteris rediviva, Doryopterissagittifolia, Doryopteris triphylla, and Doryopteris tryonii.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Dryopteris Genus selected from, but not limited to,Dryopteris abbreviata, Dryopteris acuminata, Dryopteris aemula,Dryopteris affinis, Dryopteris aitoniana, Dryopteris alpestris,Dryopteris amurensis, Dryopteris anadroma, Dryopteris antarctica,Dryopteris anthracinisquama, Dryopteris aquilinoides, Dryopterisardechensis, Dryopteris arguta, Dryopteris assimilis, Dryopterisathamantica, Dryopteris atrata, Dryopteris austriaca, Dryopterisazorica, Dryopteris barbigera, Dryopteris basisora, Dryopteris bernieri,Dryopteris bissetiana, Dryopteris bodinieri, Dryopteris borreri,Dryopteris campyloptera, Dryopteris carthusiana, Dryopteris caucasica,Dryopteris caudifrons, Dryopteris caudipinna, Dryopteris celsa,Dryopteris championii, Dryopteris chinensis, Dryopteris chrysocoma,Dryopteris cinnamomea, Dryopteris clintoniana, Dryopteris cochleata,Dryopteris commixta, Dryopteris conjugata, Dryopteris coreanomontana,Dryopteris corleyi, Dryopteris costalisora, Dryopteris crassirhizoma,Dryopteris crinalis, Dryopteris crispifolia, Dryopteris cristata,Dryopteris cycadina, Dryopteris cyclopeltidiformis, Dryopteriscystolepidota, Dryopteris decipiens, Dryopteris dehuaensis, Dryopterisdickinsii, Dryopteris diffracta, Dryopteris dilatata, Dryopteriserythrosora, Dryopteris expansa, Dryopteris fatuhivensis, Dryopterisfilix-mas, Dryopteris flaccisquama, Dryopteris formosana, Dryopterisfragrans, Dryopteris fuscipes, Dryopteris fuscoatra, Dryopteris futura,Dryopteris gamblei, Dryopteris glabra, Dryopteris goeringiana,Dryopteris goldieana, Dryopteris guanchica, Dryopteris gushanica,Dryopteris gymnophylla, Dryopteris gymnosora, Dryopteris hadanoi,Dryopteris handeliana, Dryopteris hangchowensis, Dryopteris hasseltii,Dryopteris hawaiiensis, Dryopteris hayatae, Dryopteris hendersonii,Dryopteris himachalensis, Dryopteris hondoensis, Dryopteris huberi,Dryopteris hwangii, Dryopteris inaequalis, Dryopteris indusiata,Dryopteris insularis, Dryopteris integriloba, Dryopteris intermedia,Dryopteris juxtaposita, Dryopteris karwinskyana, Dryopteris kinkiensis,Dryopteris kinokuniensis, Dryopteris knoblochii, Dryopteris koidzumiana,Dryopteris komarovii, Dryopteris labordei, Dryopteris lacera, Dryopterislachoongensis, Dryopteris laeta, Dryopteris lepidopoda, Dryopterislepidorachis, Dryopteris liankwangensis, Dryopteris ludoviciana,Dryopteris lunanensis, Dryopteris marginalis, Dryopteris marginata,Dryopteris mauiensis, Dryopteris maximowiczii, Dryopteris maxonii,Dryopteris medioxima, Dryopteris melanocarpa, Dryopteris monticola,Dryopteris munchii, Dryopteris namegatae, Dryopteris neolacera,Dryopteris nipponensis, Dryopteris nubigena, Dryopteris odontoloma,Dryopteris oligodonta, Dryopteris oreades, Dryopteris pacifica,Dryopteris pallida, Dryopteris panda, Dryopteris paraerythrosora,Dryopteris parafuscipes, Dryopteris patula, Dryopteris pentheri,Dryopteris podophylla, Dryopteris polita, Dryopteris polylepis,Dryopteris pseudofilix-mas, Dryopteris pseudosparsa, Dryopterispseudovaria, Dryopteris pulcherrima, Dryopteris pycnopteroides,Dryopteris redactopinnata, Dryopteris reflexosquamata, Dryopterisremota, Dryopteris rosea, Dryopteris rossii, Dryopteris rosthomii,Dryopteris rubiginosa, Dryopteris rubrobrunnea, Dryopteris ryo-itoana,Dryopteris sabae, Dryopteris sacrosancta, Dryopteris saffordii,Dryopteris salvinii, Dryopteris sandwicensis, Dryopteris saxifraga,Dryopteris saxifragivaria, Dryopteris scottii, Dryopteris setosa,Dryopteris shibipedis, Dryopteris shikokiana, Dryopteris shiroumensis,Dryopteris sichotensis, Dryopteris sieboldii, Dryopteris silaensis,Dryopteris simasakii, Dryopteris simplicior, Dryopteris sinofibrillosa,Dryopteris sinosparsa, Dryopteris sordidipes, Dryopteris sororia,Dryopteris sparsa, Dryopteris spinosa, Dryopteris squamifera, Dryopterissquamiseta, Dryopteris stenolepis, Dryopteris stewartii, Dryopterissubbipinnata, Dryopteris subexaltata, Dryopteris sublacera, Dryopterissubmarginata, Dryopteris submontana, Dryopteris subpycnopteroides,Dryopteris subreflexipinna, Dryopteris subtriangularis, Dryopteristetrapinnata, Dryopteris tokyoensis, Dryopteris triangularis, Dryopteristsoongii, Dryopteris tsugiwoi, Dryopteris tsutsuiana, Dryopterisunidentata, Dryopteris uniformis, Dryopteris varia, Dryopteriswallichiana, Dryopteris wattsii, Dryopteris x benedictii, Dryopterisxebinoensis, Dryopterisx triploidea, and Dryopteris yakusilvicola.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Pellaea Genus selected from, but not limited to, Pellaeaandromedifolia, Pellaea angulosa, Pellaea atropurpurea, Pellaeaboivinii, Pellaea brachyptera, Pellaea breweri, Pellaea bridgesii,Pellaea calidirupium, Pellaea calomelanos, Pellaea cordifolia, Pellaeacrenata, Pellaea cymbiformis, Pellaea doniana, Pellaea dura, Pellaeafalcata, Pellaea flavescens, Pellaea glabella, Pellaea gleichenioides,Pellaea intermedia, Pellaea longipilosa, Pellaea lyngholmii, Pellaeamaxima, Pellaea mucronata, Pellaea notabilis, Pellaea ovata, Pellaeaparadoxa, Pellaea patula, Pellaea paupercula, Pellaea pectiniformis,Pellaea pinnata, Pellaea pringlei, Pellaea pteroides, Pellaea riedelii,Pellaea rotundifolia, Pellaea rufa, Pellaea sagittata, Pellaea sp. UC1795070, Pellaea sp. UC1788706, Pellaea sp. Wen 9479, Pellaea sp. Wen9490, Pellaea ternifolia, Pellaea trichophylla, Pellaea truncata,Pellaea viridis, Pellaea wrightiana, and Pellaea glaciogena.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Gymnocarpium Genus selected from, but not limited to,Gymnocarpium appalachianum, Gymnocarpium brittonianum, Gymnocarpiumdisjunctum, Gymnocarpium Dryopteris, Gymnocarpium jessoense,Gymnocarpium oyamense, Gymnocarpium remotepinnatum, Gymnocarpiumrobertianum, and Gymnocarpium sp. TH2007-996.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Cheilanthes Genus selected from, but not limited to,Cheilanthes acrostica, Cheilanthes adiantoides, Cheilanthes aemula,Cheilanthes alabamensis, Cheilanthes austrotenuifolia, Cheilanthesbonariensis, Cheilanthes brownii, Cheilanthes catanensis, Cheilanthescaudata, Cheilanthes cavemicola, Cheilanthes clevelandii, Cheilanthescontigua, Cheilanthes cooperae, Cheilanthes covillei, Cheilanthesdistans, Cheilanthes eatonii, Cheilanthes feel, Cheilanthes fendleri,Cheilanthes fragillima, Cheilanthes glauca, Cheilanthes gracillima,Cheilanthes guanchica, Cheilanthes hispanica, Cheilanthes horridula,Cheilanthes humilis, Cheilanthes intertexta, Cheilanthesintramarginalis, Cheilanthes lanosa, Cheilanthes lasiophylla,Cheilanthes lendigera, Cheilanthes leucopoda, Cheilanthes lindheimeri,Cheilanthes maderensis, Cheilanthes microphylla, Cheilanthesmicropteris, Cheilanthes myriophylla, Cheilanthes newberryi, Cheilanthesnitida, Cheilanthes nudiuscula, Cheilanthes parryi, Cheilanthespaucijuga, Cheilanthes peninsularis, Cheilanthes persica, Cheilanthespinnatifida, Cheilanthes praetermissa, Cheilanthes prenticei,Cheilanthes pringlei, Cheilanthes pseudovellea, Cheilanthes pteroides,Cheilanthes pulchella, Cheilanthes pumilio, Cheilanthes sciadioides,Cheilanthes sieberi Kunze, Cheilanthes tenuifolia, Cheilanthes tinaei,Cheilanthes tomentosa, Cheilanthes vellea, Cheilanthes villosa,Cheilanthes viscida, Cheilanthes wootonii, Cheilanthes wrightii, andCheilanthes yavapensis.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Pteridium Genus selected from, but not limited to,Pteridium aquilinum, Pteridium arachnoideum, Pteridium brownseyi,Pteridium campestris, Pteridium capense, Pteridium caudatum, Pteridiumceheginense, Pteridium centrali-africanum, Pteridium esculentum,Pteridium falcatum, Pteridium feei, Pteridium heredia, Pteridiumlanuginosum, Pteridium latiusculum, Pteridium linea, Pteridiumpinetorum, Pteridium psittacinum, Pteridium revolutum, Pteridiumsemihastatum, Pteridium tauricum, Pteridium yarrabense, and Pteridiumyunnanense.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Christella Genus selected from, but not limited to,Christella arida, Christella augescens, Christella calvescens,Christella crinipes, Christella dentata, Christella hispidula,Christella latipinna, Christella molliuscula, Christella papilio,Christella parasitica, Christella procurrens, Christella scaberula,Christella sp. 097, Christella sp. 2257, and Christella subulata.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Lastreopsis Genus selected from, but not limited to,Lastreopsis acuminata, Lastreopsis acuta, Lastreopsis amplissima,Lastreopsis barteriana, Lastreopsis boivinii, Lastreopsis currori,Lastreopsis decomposita, Lastreopsis effusa, Lastreopsis exculta,Lastreopsis glabella, Lastreopsis hispida, Lastreopsis killipii,Lastreopsis marginans, Lastreopsis microsora, Lastreopsis munita,Lastreopsis nigritiana, Lastreopsis perrieriana, Lastreopsispseudoperrieriana, Lastreopsis rufescens, Lastreopsis silvestris,Lastreopsis smithiana, Lastreopsis sp. Kessler 1434, Lastreopsissubrecedens, Lastreopsis subsericea, Lastreopsis subsimilis, Lastreopsistenera, Lastreopsis tinarooensis, Lastreopsis vogelii, Lastreopsiswalleri, Lastreopsis windsorensis, and Lastreopsis wurunuran.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Campyloneurum Genus selected from, but not limited to,Campyloneurum abruptum, Campyloneurum aglaolepis, Campyloneurumamphostemon, Campyloneurum anetioides, Campyloneurum angustifolium,Campyloneurum angustipaleatum, Campyloneurum aphanophlebium,Campyloneurum asplundii, Campyloneurum austrobrasilianum, Campyloneurumbrevifolium, Campyloneurum centrobrasilianum, Campyloneurum chlorolepis,Campyloneurum coarctatum, Campyloneurum cochense, Campyloneurumcostatum, Campyloneurum decurrens, Campyloneurum densifolium,Campyloneurum falcoideum, Campyloneurum fasciale, Campyloneurumfuscosquamatum, Campyloneurum herbaceum, Campyloneurum inflatum,Campyloneurum lapathifolium, Campyloneurum lorentzii, Campyloneurummagnificum Moore, Campyloneurum major, Campyloneurum nitidissimum,Campyloneurum oellgaardi, Campyloneurum ophiocaulon, Campyloneurumoxypholis, Campyloneurum pascoense, Campyloneurum phyllitidis,Campyloneurum repens, Campyloneurum rigidum, Campyloneurum solutum,Campyloneurum sphenodes, Campyloneurum sublucidum, Campyloneurumtenuipes, Campyloneurum tucumanense, Campyloneurum vexatum,Campyloneurum vulpinum, Campyloneurum wacketii, Campyloneurum wurdackii,and Campyloneurum xalapense.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Hemionitis Genus selected from, but not limited to,Hemionitis acrosticha, Hemionitis acrostichoides, Hemionitisalismifolia, Hemionitis argentea, Hemionitis arifolia, Hemionitisasplenioides, Hemionitis aurea, Hemionitis aureo-nitens, Hemionitisbipinnata, Hemionitis blumeana, Hemionitis boryanum, Hemionitisbrasiliana, Hemionitis cajenensis, Hemionitis callifolia, Hemionitischaerophylla, Hemionitis citrifolia, Hemionitis concava, Hemionitiscordata, Hemionitis cordifolia, Hemionitis coriacea, Hemionitiscumingiana, Hemionitis dealbata, Hemionitis discolor, Hemionitiselegans, Hemionitis elongata, Hemionitis esculenta, Hemionitis falcata,Hemionitis gigantea, Hemionitis grandifolia, Hemionitis griffithii,Hemionitis gymnopteroidea, Hemionitis hastata, Hemionitis hederifolia,Hemionitis hookeriana, Hemionitis hosei, Hemionitis humilis, Hemionitisimmersa, Hemionitis incisa, Hemionitis intermedia, Hemionitis japonica,Hemionitis lanceolata, Hemionitis latifolia, Hemionitis leptophylla,Hemionitis lessonii, Hemionitis levyi, Hemionitis lineata, Hemionitismaingayi, Hemionitis muelleri, Hemionitis obtusa, Hemionitis opaca,Hemionitis otonis, Hemionitis palmata, Hemionitis parasitica, Hemionitisparvula, Hemionitis pedata, Hemionitis pedatifida, Hemionitis pinnata,Hemionitis pinnatifida, Hemionitis plantaginea, Hemionitis podophylla,Hemionitis polypodioides, Hemionitis pothifolia, Hemionitis pozoi,Hemionitis prolifera, Hemionitis reinwardtiana, Hemionitis reticulata,Hemionitis rigida, Hemionitis rufa, Hemionitis sagittata, Hemionitissemicostata, Hemionitis sessilifolia, Hemionitisx smithii, Hemionitisspatulata, Hemionitis stipitata, Hemionitis subcordata, Hemionitistomentosa, Hemionitis toxotis, Hemionitis triloba, Hemionitis trinervis,Hemionitis vestita, Hemionitis vittaeformis, Hemionitis wilfordii, andHemionitis zollingeri.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Selliguea Genus selected from, but not limited to,Selliguea albicaula, Selliguea albidopaleata, Selliguea albidosquamata,Selliguea albopes, Selliguea archboldii, Selliguea bakeri, Selligueabalbi, Selliguea banaensis, Selliguea bellisquamata, Selligueabisulcata, Selliguea brooksii, Selliguea caudiformis, Selligueaceratophylla, Selliguea chenkouensis, Selliguea chinensis, Selligueachrysotricha, Selliguea conjuncta, Selliguea connexa, Selligueacostulata, Selliguea craspedosora, Selliguea crenatopinnata, Selligueacretifera, Selliguea cruciformis, Selliguea cunea, Selliguea dactylina,Selliguea dekockii, Selliguea digitata, Selliguea ebenipes, Selligueaechinospora, Selliguea elmeri, Selliguea enervis, Selliguea engleri,Selliguea erythrocarpa, Selliguea feel, Selliguea feeoides, Selligueaferrea, Selliguea fukienensis, Selliguea glauca, Selliguea glaucopsis,Selliguea gracilipes, Selliguea griffithiana, Selliguea hainanensis,Selliguea hastata, Selliguea hellwigii, Selliguea heterocarpa, Selligueahirsuta, Selliguea hirtella, Selliguea hunyaensis, Selligueaintegerrima, Selliguea katuii, Selliguea kingpingensis, Selligueakwangtungensis, Selliguea laciniata, Selliguea lagunensis, Selliguealaipoensis, Selliguea lancea, Selliguea lanceola, Selliguea lateritia,Selliguea lauterbachii, Selliguea likiangensis, Selliguea majoensis,Selliguea malacodon, Selliguea metacoela, Selliguea montana, Selligueamurudensis, Selliguea neglecta, Selliguea nigropaleacea, Selligueanigrovenia, Selliguea oblongifolia, Selliguea obtusa, Selligueaomeiensis, Selliguea oodes, Selliguea oxyloba, Selliguea palmatifida,Selliguea pampylocarpa, Selliguea pellucidifolia, Selliguea pianmaensis,Selliguea pingpienensis, Selliguea plantaginea, Selliguea platyphylla,Selliguea pseudoacrosticha, Selliguea pyrolifolia, Selligueaquasidivaricata, Selliguea rhynchophylla, Selliguea rigida, Selliguearoseomarginata, Selliguea rotunda, Selliguea setacea, Selligueashandongensis, Selliguea shensiensis, Selliguea similis, Selligueasimplicifolia, Selliguea simplicissima, Selliguea soridens, Selligueasri-ratu, Selliguea stenophylla, Selliguea stenosquamis, Selligueastewartii, Selliguea suboxyloba, Selliguea subsparsa, Selligueasubtaeniata, Selliguea taeniata, Selliguea tafana, Selligueataiwanensis, Selliguea tamdaoensis, Selliguea tarningensis, Selligueatenuipes, Selliguea tibetana, Selliguea triloba, Selliguea triquetra,Selliguea violascens, Selliguea waltonii, Selliguea whitfordii,Selliguea wuliangshanensis, Selliguea wuyishanica, Selligueayakuinsularis, and Selliguea yakushimensis.

In some embodiments, the IPD113 polypeptide is derived from a fernspecies in the Arachniodes Genus selected from, but not limited to,Arachniodes abrupta, Arachniodes acuminata, Arachniodes ailaoshanensis,Arachniodes amabilis, Arachniodes amoena, Arachniodes anshunensis,Arachniodes argillicola, Arachniodes arisanica, Arachniodes aristata,Arachniodes aristatissima, Arachniodes aspidioides, Arachniodesassamica, Arachniodes attenuata, Arachniodes australis, Arachniodesaustro-yunnanensis, Arachniodes x azuminoensis, Arachniodes baiseensis,Arachniodes basipinnata, Arachniodes bella, Arachniodes bipinnata,Arachniodes blinii, Arachniodes borealis, Arachniodes calcarata,Arachniodes cantilenae, Arachniodes carvifolia, Arachniodes caudata,Arachniodes caudifolia, Arachniodes cavalerii, Arachniodescentrochinensis, Arachniodes chaerophylloides, Arachniodes chinensis,Arachniodes ii, Arachniodes clivorum, Arachniodes coadnata, Arachniodesconiifolia, Arachniodes cornopteris, Arachniodes cornucervi, Arachniodescostulisora, Arachniodes cyrtomifolia, Arachniodes damiaoshanensis,Arachniodes davalliaeformis, Arachniodes dayaoensis, Arachniodesdecomposita, Arachniodes denticulata, Arachniodes denticulata,Arachniodes denticulatabarbensis, Arachniodes denticulatajucunda,Arachniodes diffracta, Arachniodes dimorphophyllum, Arachniodesduplicatoserrata, Arachniodes elevatas, Arachniodes emeiensis,Arachniodes erythrosora, Arachniodes exilis, Arachniodes falcata,Arachniodes fengii, Arachniodes fengyangshanensis, Arachniodes festina,Arachniodes foeniculacea, Arachniodes foliosa, Arachniodes formosa,Arachniodes formosissima, Arachniodes fujianensis, Arachniodesfuteshanensis, Arachniodes gansuensis, Arachniodes gigantea, Arachniodesgijiangensis, Arachniodes gizushanensis, Arachniodes globisora,Arachniodes gongshanensis, Arachniodes gradata, Arachniodes grossa,Arachniodes guangnanensis, Arachniodes guangtongensis, Arachniodesguangxiensis, Arachniodes guanxianensis, Arachniodes hainanensis,Arachniodes haniffii, Arachniodes hasseltii, Arachniodes hekiana,Arachniodes hekouensis, Arachniodes henryi, Arachniodes heyuanensis,Arachniodes hiugana, Arachniodes holttumii, Arachniodes huapingensis,Arachniodes hunanensis, Arachniodes hupingshanensis, Arachniodesxikeminensis, Arachniodes insularis, Arachniodes intermedia, Arachniodesishingensis, Arachniodes japonica, Arachniodes jiangxiensis, Arachniodesjinfoshanensis, Arachniodes jingdongensis, Arachniodes jinpingensis,Arachniodes jiulongshanensis, Arachniodes kansuensis, Arachniodeskenzo-satakei, Arachniodes kurosawae, Arachniodes kweichowensis,Arachniodes lanceolata, Arachniodes leuconeura, Arachniodesleucostegioides, Arachniodes liyangensis, Arachniodes longipinna,Arachniodes lurida, Arachniodes lushanensis, Arachniodes lushuiensis,Arachniodes macrocarpa, Arachniodes macrostegia, Arachniodesmacrostegia, Arachniodes maguanensis, Arachniodes maoshanensis,Arachniodes masakii, Arachniodes maxima, Arachniodes maximowiczii,Arachniodes maximowiczii, Arachniodes menglianensis, Arachniodesmengziensis, Arachniodes michelii, Arachniodes minamitanii, Arachniodesmiqueliana, Arachniodes mirabilis, Arachniodesx mitsuyoshiana,Arachniodes multifida, Arachniodes mutica, Arachniodes nanchuanensis,Arachniodes nanqingensis, Arachniodes neoaristata, Arachniodesneobipinnata, Arachniodes neofalcata, Arachniodes neohunanensis,Arachniodes neopodophylla, Arachniodes nibashanensis, Arachniodesnigrospinosa, Arachniodes nipponica, Arachniodes nitidula, Arachniodesobtusiloba, Arachniodes obtusipinnula, Arachniodes obtusissima,Arachniodes ochropteroides, Arachniodes okinawensis, Arachniodesoohorae, Arachniodes palmipes, Arachniodes parasimplicior, Arachniodespianmaensis, Arachniodes pinnatifida, Arachniodes pseudo-assamica,Arachniodes pseudo-longipinna, Arachniodes pseudo-repens, Arachniodespseudo-simplicior, Arachniodes pseudoaristata, Arachniodespseudocavalerii, Arachniodes x pseudohekiana, Arachniodes pubescens,Arachniodes puncticulata, Arachniodes quadripinnata, Arachniodesreducta, Arachniodes repens, Arachniodes respiciens, Arachniodes xrespiciens, Arachniodes rhomboidea, Arachniodes rhomboidearhomboidea,Arachniodes rigidissima, Arachniodes sarasiniorum, Arachniodessasamotoi, Arachniodes semifertilis, Arachniodes setifera, Arachniodesshuangbaiensis, Arachniodes sichuanensis, Arachniodes similis,Arachniodes simplicior, Arachniodes simulans, Arachniodes sino-aristata,Arachniodes sino-rhomboidea, Arachniodes sinomiqueliana, Arachniodessledgei, Arachniodes sparsa, Arachniodes speciosa, Arachniodesspectabilis, Arachniodes sphaerosora, Arachniodes spino-serrulata,Arachniodes sporadosora, Arachniodes squamulosa, Arachniodes standishii,Arachniodes subamabilis, Arachniodes subamoena, Arachniodes subaristata,Arachniodes subreflexipinna, Arachniodes suijiangensis, Arachniodessuperba, Arachniodes x takayamensis, Arachniodes tibetana, Arachniodestiendongensis, Arachniodes tomitae, Arachniodes tonkinensis, Arachniodestriangularis, Arachniodes tripinnata, Arachniodes tsiangiana,Arachniodes valida, Arachniodes walkerae, Arachniodes webbiana,Arachniodes wulingshanensis, Arachniodes xinpingensis, Arachniodesyakusimensis, Arachniodes yandangshanensis, Arachniodes yaomashanensis,Arachniodes yaoshanensis, Arachniodes yasu-inouei, Arachniodesyinjiangensis, Arachniodes yixinensis, Arachniodes yoshinagae,Arachniodes yunnanensis, Arachniodes yunqiensis, Arachniodes zeylanica,and Arachniodes ziyunshanensis.

“Sufficiently homologous” is used herein to refer to an amino acidsequence that has at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 6%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or greater sequence homology compared to a reference sequenceusing one of the alignment programs described herein using standardparameters. In some embodiments, the sequence homology is against thefull-length sequence of an IPD113 polypeptide.

In some embodiments, the IPD113 polypeptide has at least about 40%, 45%,50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 8%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identitycompared to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ IDNO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19,SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO:24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ IDNO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38,SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO:43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ IDNO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57,SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO:62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ IDNO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76,SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO:81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ IDNO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95,SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO:118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO:253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO:262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO:271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO:280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO:318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO:425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO:434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO:447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO:463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO:478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO:487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQID NO: 492, SEQ ID NO: 494, and SEQ ID NO: 495, as well as amino acidsubstitutions, deletions, insertions, fragments thereof, andcombinations thereof.

In some embodiments, the IPD113 polypeptide has at least about 40%, 45%,50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identitycompared to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ IDNO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19,SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO:24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ IDNO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38,SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO:43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ IDNO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57,SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO:62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ IDNO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76,SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO:81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ IDNO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95,SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO:118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO:416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO:428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO:438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO:451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO:470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO:481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO:490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494, and SEQ ID NO: 495and has at least one amino acid substitution, deletion, insertion orcombination therefore compared to the native sequence.

In another aspect IPD113 polypeptides are encompassed. Also provided areisolated or recombinant IPD113 polypeptides of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7,SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12,SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ IDNO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31,SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO:36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ IDNO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO:55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ IDNO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69,SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO:74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ IDNO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88,SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO:93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ IDNO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102,SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ IDNO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111,SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ IDNO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120,SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ IDNO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255,SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ IDNO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264,SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ IDNO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273,SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ IDNO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311,SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ IDNO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320,SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ IDNO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427,SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ IDNO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436,SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ IDNO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450,SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ IDNO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469,SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ IDNO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480,SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ IDNO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489,SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494, or SEQID NO: 495. The term “about” when used herein in context with percentsequence identity means +/−0.5%. One of skill in the art will recognizethat these values can be appropriately adjusted to determinecorresponding homology of proteins considering amino acid similarity andthe like.

In some embodiments, the sequence identity is calculated using ClustalWalgorithm in the ALIGNX® module of the Vector NTI® Program Suite(Invitrogen Corporation, Carlsbad, Calif.) with all default parameters.In some embodiments, the sequence identity is across the entire lengthof polypeptide calculated using ClustalW algorithm in the ALIGNX® moduleof the Vector NTI® Program Suite (Invitrogen Corporation, Carlsbad,Calif.) with all default parameters.

As used herein, the terms “protein,” “peptide molecule,” or“polypeptide” includes any molecule that comprises five or more aminoacids. Protein, peptide or polypeptide molecules may undergomodification, including post-translational modifications, such as, butnot limited to, disulfide bond formation, glycosylation, phosphorylationor oligomerization. Thus, as used herein, the terms “protein,” “peptidemolecule” or “polypeptide” includes any protein that is modified by anybiological or non-biological process. The terms “amino acid” and “aminoacids” refer to all naturally occurring L-amino acids.

A “recombinant protein” is used herein to refer to a protein that is nolonger in its natural environment, for example in vitro or in arecombinant bacterial or plant host cell. An IPD113 polypeptide that issubstantially free of cellular material includes preparations of proteinhaving less than about 30%, 20%, 10% or 5% (by dry weight) ofnon-pesticidal protein (also referred to herein as a “contaminatingprotein”).

“Fragments” or “biologically active portions” include polypeptidefragments comprising amino acid sequences sufficiently identical to anIPD113 polypeptide and that exhibit insecticidal activity. “Fragments”or “biologically active portions” of IPD113 polypeptides includesfragments comprising amino acid sequences sufficiently identical to theamino acid sequence set forth in IPD113 polypeptides of SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6,SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11,SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ IDNO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30,SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO:35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ IDNO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO:54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ IDNO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68,SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO:73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ IDNO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO:92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ IDNO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ IDNO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110,SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ IDNO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119,SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ IDNO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254,SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ IDNO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263,SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ IDNO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272,SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ IDNO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281,SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ IDNO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319,SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ IDNO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426,SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ IDNO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435,SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ IDNO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448,SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ IDNO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466,SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ IDNO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479,SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ IDNO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488,SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ IDNO: 494, or SEQ ID NO: 495, wherein the polypeptide has insecticidalactivity. Such biologically active portions can be prepared byrecombinant techniques and evaluated for insecticidal activity. In someembodiments, the IPD113 polypeptide fragment is an N-terminal and/or aC-terminal truncation of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31 or more amino acids from the N-terminus and/or C-terminus relative toIPD113 polypeptides of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 32, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495, e.g., byproteolysis, by insertion of a start codon, by deletion of the codonsencoding the deleted amino acids and concomitant insertion of a startcodon, and/or insertion of a stop codon.

In some embodiments, the IPD113 polypeptide fragment is an N-terminaltruncation of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24 amino acids from the N-terminusof IPD113 polypeptides of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID , SEQID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO:257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO:266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO:275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO:313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO:419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO:429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO:439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO:452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO:472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO:482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO:491, SEQ ID NO: 492, SEQ ID NO: 494, or SEQ ID NO: 495.

In some embodiments, the IPD113 polypeptide fragment is an N-terminaland/or a C-terminal truncation of at least 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, 30, 31, 32, 33, 34 or more amino acids from the N-terminusand/or C-terminus relative to IPD113 polypeptides of SEQ ID NO: 1, SEQID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO:255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO:264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO:273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO:311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO:320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO:427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO:436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO:450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO:469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO:480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO:489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 orSEQ ID NO: 495.

“Variants” as used herein refers to proteins or polypeptides having anamino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or greater identical to the parental aminoacid sequence.

In some embodiments, an IPD113 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or greater identity to the amino acid sequence of IPD113polypeptides of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO:33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ IDNO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47,SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO:52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ IDNO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66,SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO:71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ IDNO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85,SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO:90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ IDNO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO:104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO:113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO:122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO:257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO:266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO:275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO:313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO:419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO:429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO:439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO:452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO:472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO:482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO:491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495, wherein theIPD113 polypeptide has insecticidal activity.

In some embodiments, an IPD113 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greateridentity across the entire length of the amino acid sequence of theIPD113 polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID

NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90,SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO:95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ IDNO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104,SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ IDNO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113,SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ IDNO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122,SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ IDNO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257,SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ IDNO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266,SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ IDNO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275,SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ IDNO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313,SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ IDNO: 318, SEQ ID NO: 319 or SEQ ID NO: 320.

In some embodiments, the sequence identity is across the entire lengthof the polypeptide calculated using ClustalW algorithm in the ALIGNX®module of the Vector NTI® Program Suite (Invitrogen Corporation,Carlsbad, Calif.) with all default parameters.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ IDNO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 27, SEQ ID NO: 30,SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO:77, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ IDNO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQID NO: 98, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO:103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113 or SEQ ID NO:114.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 24, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO:95, SEQ ID NO: 96, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQID NO: 106, SEQ ID NO: 107, SEQ ID NO: 39 or SEQ ID NO: 40.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90 or SEQ IDNO: 91.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 20, SEQ ID NO: 24 or SEQ ID NO: 27.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:21 or SEQ ID NO: 22.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO:38, SEQ ID NO: 77, SEQ ID NO: 110, SEQ ID NO: 111 or SEQ ID NO: 112.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater identity across the entire length of the aminoacid sequence of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO: 16, SEQ ID NO: 23, SEQ ID NO: 35, SEQ ID NO: 36, SEQ IDNO: 37, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 113 or SEQ ID NO: 114.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greateridentity across the entire length of the amino acid sequence of SEQ IDNO: 10.

In some embodiments, the IPD113 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greateridentity across the entire length of the amino acid sequence of SEQ IDNO: 16.

In some embodiments, an IPD113 polypeptide comprises an amino acidsequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ IDNO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19,SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO:24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ IDNO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38,SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO:43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ IDNO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57,SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO:62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ IDNO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76,SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO:81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ IDNO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95,SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO:118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO:253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO:262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO:271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO:280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO:318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO:425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO:434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO:447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO:463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO:478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO:487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 having 1, 2, 3, 4, 5, 6, 7,8, 9, 10 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35 or more amino acid substitutions,deletions and/or insertions compared to the native amino acid at thecorresponding position of IPD113 polypeptides of SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123,

SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ IDNO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258,SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ IDNO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267,SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ IDNO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276,SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ IDNO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314,SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ IDNO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420,SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ IDNO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430,SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ IDNO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440,SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ IDNO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453,SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ IDNO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473,SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ IDNO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483,SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ IDNO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492,SEQ ID NO: 494 or SEQ ID NO: 495.

Methods for such manipulations of an IPD113 polypeptide can be preparedby mutations in the DNA. This may also be accomplished by one of severalforms of mutagenesis and/or in directed evolution.

In some aspects, the changes encoded in the amino acid sequence will notsubstantially affect the function of the protein. Such variants willpossess the desired pesticidal activity. However, it is understood thatthe ability of an IPD113 polypeptide to confer pesticidal activity maybe improved using such techniques upon the compositions of thisdisclosure.

For example, conservative amino acid substitutions may be made at one ormore predicted nonessential amino acid residues. A “nonessential” aminoacid residue is a residue that can be altered from the wild-typesequence of an IPD113 polypeptide without altering the biologicalactivity. Alignment of the amino acid sequences of IPD113 polypeptidehomologs (for example—FIGS. 2, 3, and 4 ), allows for the identificationof residues that are highly conserved amongst the natural homologs ofthis family as well as residues or regions tolerant to amino aciddiversity. A “conservative amino acid substitution” is one in which theamino acid residue is replaced with an amino acid residue having asimilar side chain. Families of amino acid residues having similar sidechains have been defined in the art. These families include: amino acidswith basic side chains (e.g., lysine, arginine, histidine); acidic sidechains (e.g., aspartic acid, glutamic acid); polar, negatively chargedresidues and their amides (e.g., aspartic acid, asparagine, glutamic,acid, glutamine; uncharged polar side chains (e.g., glycine, asparagine,glutamine, serine, threonine, tyrosine, cysteine); small aliphatic,nonpolar or slightly polar residues (e.g., Alanine, serine, threonine,proline, glycine); nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine, tryptophan); largealiphatic, nonpolar residues (e.g., methionine, leucine, isoleucine,valine, cystine); beta-branched side chains (e.g., threonine, valine,isoleucine); aromatic side chains (e.g., tyrosine, phenylalanine,tryptophan, histidine); large aromatic side chains (e.g., tyrosine,phenylalanine, tryptophan).

Amino acid substitutions may be made in nonconserved regions that retainfunction. In general, such substitutions would not be made for conservedamino acid residues or for amino acid residues residing within aconserved motif, where such residues are essential for protein activity.Examples of residues that are conserved and that may be essential forprotein activity include, for example, residues that are identicalbetween all proteins contained in an alignment of similar or relatedtoxins to the sequences of the embodiments (e.g., residues that areidentical in an alignment of homologous proteins). Examples of residuesthat are conserved but that may allow conservative amino acidsubstitutions and still retain activity include, for example, residuesthat have only conservative substitutions between all proteins containedin an alignment of similar or related toxins to the sequences of theembodiments (e.g., residues that have only conservative substitutionsbetween all proteins contained in the alignment homologous proteins).However, one of skill in the art would understand that functionalvariants may have minor conserved or nonconserved alterations in theconserved residues. Guidance as to appropriate amino acid substitutionsthat do not affect biological activity of the protein of interest may befound in the model of Dayhoff, et al., (1978) Atlas of Protein Sequenceand Structure (Natl. Biomed. Res. Found., Washington, D.C.), hereinincorporated by reference.

In making such changes, the hydropathic index of amino acids may beconsidered. The importance of the hydropathic amino acid index inconferring interactive biologic function on a protein is generallyunderstood in the art (Kyte and Doolittle, (1982) J Mol Biol.157(1):105-32).

It is accepted that the relative hydropathic character of the amino acidcontributes to the secondary structure of the resultant protein, whichin turn defines the interaction of the protein with other molecules, forexample, enzymes, substrates, receptors, DNA, antibodies, antigens, andthe like.

Certain amino acids may be substituted by other amino acids having asimilar hydropathic index or score and still result in a protein withsimilar biological activity, i.e., still obtain a biologicalfunctionally equivalent protein. Each amino acid has been assigned ahydropathic index based on its hydrophobicity and charge characteristics(Kyte and Doolittle, ibid). These are: isoleucine (+4.5); valine (+4.2);leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5);methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (−0.7);serine (−0.8); tryptophan (−0.9); tyrosine (−1.3); proline (−1.6);histidine (−3.2); glutamate (−3.5); glutamine (−3.5); aspartate (−3.5);asparagine (−3.5); lysine (−3.9) and arginine (−4.5).

In making such changes, the substitution of amino acids whosehydropathic indices are within +2 is preferred, those which are within+1 are particularly preferred, and those within +0.5 are even moreparticularly preferred.

It is also understood that the substitution of like amino acids can bemade effectively based on hydrophilicity. U.S. Pat. No. 4,554,101,states that the greatest local average hydrophilicity of a protein, asgoverned by the hydrophilicity of its adjacent amino acids, correlateswith a biological property of the protein.

As detailed in U.S. Pat. No. 4,554,101, the following hydrophilicityvalues have been assigned to amino acid residues: arginine (+3.0);lysine (+3.0); aspartate (+3.0.+0.1); glutamate (+3.0.+0.1); serine(+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine(−0.4); proline (−0.5.+0.1); alanine (−0.5); histidine (−0.5); cysteine(−1.0); methionine (−1.3); valine (−1.5); leucine (−1.8); isoleucine(−1.8); tyrosine (−2.3); phenylalanine (−2.5); tryptophan (−3.4).

Alternatively, alterations may be made to the protein sequence of manyproteins at the amino or carboxy terminus without substantiallyaffecting activity. This can include insertions, deletions oralterations introduced by modern molecular methods, such as PCR,including PCR amplifications that alter or extend the protein codingsequence by inclusion of amino acid encoding sequences in theoligonucleotides utilized in the PCR amplification. Alternatively, theprotein sequences added can include entire protein-coding sequences,such as those used commonly in the art to generate protein fusions. Suchfusion proteins are often used to (1) increase expression of a proteinof interest (2) introduce a binding domain, enzymatic activity orepitope to facilitate either protein purification, protein detection orother experimental uses (3) target secretion or translation of a proteinto a subcellular organelle, such as the periplasmic space ofGram-negative bacteria, mitochondria or chloroplasts of plants or theendoplasmic reticulum of eukaryotic cells, the latter of which oftenresults in glycosylation of the protein.

Variant nucleotide and amino acid sequences of the disclosure alsoencompass sequences derived from mutagenic and recombinogenic proceduressuch as DNA shuffling. With such a procedure, one or more differentIPD113 polypeptide coding regions can be used to create a new IPD113polypeptide possessing the desired properties. In this manner, librariesof recombinant polynucleotides are generated from a population ofrelated sequence polynucleotides comprising sequence regions that havesubstantial sequence identity and can be homologously recombined invitro or in vivo. For example, using this approach, sequence motifsencoding a domain of interest may be shuffled between a pesticidal geneand other known pesticidal genes to obtain a new gene coding for aprotein with an improved property of interest, such as an increasedinsecticidal activity. Strategies for such DNA shuffling includeStemmer, (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751; Stemmer,(1994) Nature 370:389-391; Crameri, et al., (1997) Nature Biotech.15:436-438; Moore, et al., (1997) J. Mol. Biol. 272:336-347; Zhang, etal., (1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Crameri, et al.,(1998) Nature 391:288-291; and U.S. Pat. Nos. 5,605,793 and 5,837,458.

Domain swapping or shuffling is another mechanism for generating alteredIPD113 polypeptides. Domains may be swapped between IPD113 polypeptidesresulting in hybrid or chimeric toxins with improved insecticidalactivity or target spectrum. Methods for generating recombinant proteinsand testing them for pesticidal activity (see, for example, Naimov, etal., (2001) Appl. Environ. Microbiol. 67:5328-5330; de Maagd, et al.,(1996) Appl. Environ. Microbiol. 62:1537-1543; Ge, et al., (1991) J.Biol. Chem. 266:17954-17958; Schnepf, et al., (1990) J. Biol. Chem.265:20923-20930; Rang, et al., 91999) Appl. Environ. Microbiol.65:2918-2925).

Phylogenetic, sequence motif, and structural analyses of insecticidalprotein families. A sequence and structure analysis method can beemployed, which is composed of four components: phylogenetic treeconstruction, protein sequence motifs finding, secondary structureprediction, and alignment of protein sequences and secondary structures.Details about each component are illustrated below.

1) Phylogenetic Tree Construction

The phylogenetic analysis can be performed using the software MEGAS.Protein sequences can be subjected to ClustalW version 2 analysis(Larkin M. A et al (2007) Bioinformatics 23(21): 2947-2948) for multiplesequence alignment. The evolutionary history is then inferred by theMaximum Likelihood method based on the JTT matrix-based model. The treewith the highest log likelihood is obtained, exported in Newick format,and further processed to extract the sequence IDs in the same order asthey appeared in the tree. A few clades representing sub-families can bemanually identified for each insecticidal protein family.

2) Protein Sequence Motifs Finding

Protein sequences are re-ordered according to the phylogenetic treebuilt previously, and fed to the MOTIF analysis tool MEME (Multiple EMfor MOTIF Elicitation) (Bailey T. L., and Elkan C., Proceedings of theSecond International Conference on Intelligent Systems for MolecularBiology, pp. 28-36, AAAI Press, Menlo Park, California, 1994.) foridentification of key sequence motifs. MEME is setup as follows: Minimumnumber of sites 2, Minimum motif width 5, and Maximum number of motifs30. Sequence motifs unique to each sub-family were identified by visualobservation. The distribution of MOTIFs across the entire gene familycould be visualized in HTML webpage. The MOTIFs are numbered relative tothe ranking of the E-value for each MOTIF.

3) Secondary Structure Prediction

PSIPRED, top ranked secondary structure prediction method (Jones DT.(1999) J. Mol. Biol. 292: 195-202), can be used for protein secondarystructure prediction. The tool provides accurate structure predictionusing two feed-forward neural networks based on the PSI-BLAST output.The PSI-BLAST database is created by removing low-complexity,transmembrane, and coiled-coil regions in Uniref100. The PSIPRED resultscontain the predicted secondary structures (Alpha helix: H, Beta strand:E, and Coil: C) and the corresponding confidence scores for each aminoacid in a given protein sequence.

4) Alignment of Protein Sequences and Secondary Structures

A script can be developed to generate gapped secondary structurealignment according to the multiple protein sequence alignment from step1 for all proteins. All aligned protein sequences and structures areconcatenated into a single FASTA file, and then imported into MEGA forvisualization and identification of conserved structures.

In some embodiments, the IPD113 polypeptide has a modified physicalproperty. As used herein, the term “physical property” refers to anyparameter suitable for describing the physical-chemical characteristicsof a protein. As used herein, “physical property of interest” and“property of interest” are used interchangeably to refer to physicalproperties of proteins that are being investigated and/or modified.Examples of physical properties include, but are not limited to, netsurface charge and charge distribution on the protein surface, nethydrophobicity and hydrophobic residue distribution on the proteinsurface, surface charge density, surface hydrophobicity density, totalcount of surface ionizable groups, surface tension, protein size and itsdistribution in solution, melting temperature, heat capacity, and secondvirial coefficient. Examples of physical properties also include, IPD113polypeptide having increased expression, increased solubility, decreasedphytotoxicity, and digestibility of proteolytic fragments in an insectgut. Models for digestion by simulated gastric fluids include those ofFuchs, R. L. and J. D. Astwood. Food Technology 50: 83-88, 1996;Astwood, J. D., et al Nature Biotechnology 14: 1269-1273, 1996; Fu T Jet al J. Agric Food Chem. 50: 7154-7160, 2002.

In some embodiments, variants include polypeptides that differ in aminoacid sequence due to mutagenesis. Variant proteins encompassed by thedisclosure are biologically active, that is they continue to possess thedesired biological activity (i.e. pesticidal activity) of the nativeprotein. In some embodiment, the variant will have at least about 10%,at least about 30%, at least about 50%, at least about 70%, at leastabout 80% or more of the insecticidal activity of the native protein. Insome embodiments, the variants may have improved activity over thenative protein.

Bacterial genes quite often possess multiple methionine initiationcodons in proximity to the start of the open reading frame. Often,translation initiation at one or more of these start codons will lead togeneration of a functional protein. These start codons can include ATGcodons. However, bacteria such as Bacillus sp. also recognize the codonGTG as a start codon, and proteins that initiate translation at GTGcodons contain a methionine at the first amino acid. On rare occasions,translation in bacterial systems can initiate at a TTG codon, though inthis event the TTG encodes a methionine. Furthermore, it is not oftendetermined a priori which of these codons are used naturally in thebacterium. Thus, it is understood that use of one of the alternatemethionine codons may also lead to generation of pesticidal proteins.These pesticidal proteins are encompassed in the present disclosure andmay be used in the methods of the present disclosure. It will beunderstood that, when expressed in plants, it will be necessary to alterthe alternate start codon to ATG for proper translation.

One skilled in the art understands that the polynucleotide codingsequence can be modified to add a codon at the penultimate positionfollowing the methionine start codon to create a restriction enzyme sitefor recombinant cloning purposes and/or for expression purposes. In someembodiments, the IPD113 polypeptide further comprises an alanine residueat the position after the translation initiator methionine.

In some embodiments, the translation initiator methionine of the IPD113polypeptide is cleaved off post translationally. One skilled in the artunderstands that the N-terminal translation initiator methionine can beremoved by methionine aminopeptidase in many cellular expressionsystems.

In some embodiments, the IPD113 polypeptide comprises the amino acidsequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ IDNO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19,SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO:24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ IDNO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38,SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO:43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ IDNO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57,SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO:62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ IDNO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76,SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO:81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ IDNO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95,SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO:118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO:253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO:262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO:271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO:280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO:318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO:425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO:434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO:447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO:463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO:478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO:487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495.

In some embodiments, chimeric polypeptides are provided comprisingregions of at least two different IPD113 polypeptides of the disclosure.

In some embodiments, chimeric polypeptides are provided comprisingregions of at least two different IPD113 polypeptides selected from SEQID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ IDNO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ IDNO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20,SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ IDNO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39,SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO:44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ IDNO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58,SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO:63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ IDNO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77,SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO:82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ IDNO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96,SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO:101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO:110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO:119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO:254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO:263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO:272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO:281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO:319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO:426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO:435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO:448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO:466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO:479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID

NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492,SEQ ID NO: 494 or SEQ ID NO: 495.

In some embodiments, the chimeric insecticidal protein comprises a) afirst portion of a continuous stretch of the amino acid sequence of afirst recombinant IPD113 polypeptide of the embodiments; and b) acorresponding second portion of a continuous stretch of the amino acidsequence of a second recombinant IPD113 polypeptide of the embodiments.As used herein “corresponding portion” means a part or all the aminoacid sequence of the second recombinant IPD113 polypeptide thatcontinues from the end or breakpoint of the portion of the amino acidsequence of the first recombinant IPD113 polypeptide. As used herein,“breakpoint” means the transition point between the two IPD113 popeptidesequences. For example, if the first and second IPD113 polypeptide areabout three hundred amino acids in length and the first portion includesamino acids 1 to about 175 from the first IPD113 polypeptide and thecorresponding second portion comprises amino acids about amino acids 176to about 300 from the second IPD113 polypeptide. It is understood thatthe two sequences may vary in length so the residue numbering may not bethe same between the two sequences. An amino acid sequence alignmentbetween two or more IPD113 polypeptides can be used to determine thecorrespondence between the numbering of the residues of the polypeptidesequences. A breakpont can be selected based on but not limited to: 1)shared regions of sequence between the two IPD113 polypeptides; 2)between shared domains or motifs; or 3) between shared secondary ortertiary structural elements. The occurrence of sequences within regionsthat are known to be involved in periodic secondary structure (alpha and3-10 helices, parallel and anti-parallel beta sheets) are regions thatshould be avoided. Similarly, regions of amino acid sequence that areobserved or predicted to have a low degree of solvent exposure are morelikely to be part of the so-called hydrophobic core of the protein andshould also be avoided for selection of the breakpoint. In contrast,those regions that are known or predicted to be in surface turns orloops, and especially those regions that are known not to be requiredfor biological activity, are the preferred sites for location of thebreakpoint. Continuous stretches of amino acid sequence that arepreferred based on the above criteria are referred to as a “breakpointregion”.

In some embodiments, chimeric IPD113 polypeptide are provided comprisingan N-terminal Region of a first IPD113 polypeptide of the disclosureoperably fused to a C-terminal Region of a second IPD113 polypeptide ofthe disclosure.

In other embodiments, the IPD113 polypeptide may be expressed as aprecursor protein with an intervening sequence that catalyzesmulti-step, post translational protein splicing. Protein splicinginvolves the excision of an intervening sequence from a polypeptide withthe concomitant joining of the flanking sequences to yield a newpolypeptide (Chong, et al., (1996) J. Biol. Chem., 271:22159-22168).This intervening sequence or protein splicing element, referred to asinteins, which catalyze their own excision through three coordinatedreactions at the N-terminal and C-terminal splice junctions: an acylrearrangement of the N-terminal cysteine or serine; a transesterficationreaction between the two termini to form a branched ester or thioesterintermediate and peptide bond cleavage coupled to cyclization of theintein C-terminal asparagine to free the intein (Evans, et al., (2000)J. Biol. Chem., 275:9091-9094. The elucidation of the mechanism ofprotein splicing has led to a number of intein-based applications (Comb,et al., U.S. Pat. No. 5,496,714; Comb, et al., U.S. Pat. No. 5,834,247;Camarero and Muir, (1999) J. Amer. Chem. Soc. 121:5597-5598; Chong, etal., (1997) Gene 192:271-281, Chong, et al., (1998) Nucleic Acids Res.26:5109-5115; Chong, et al., (1998) J. Biol. Chem. 273:10567-10577;Cotton, et al., (1999) J. Am. Chem. Soc. 121:1100-1101; Evans, et al.,(1999) J. Biol. Chem. 274:18359-18363; Evans, et al., (1999) J. Biol.Chem. 274:3923-3926; Evans, et al., (1998) Protein Sci. 7:2256-2264;Evans, et al., (2000) J. Biol. Chem. 275:9091-9094; Iwai and Pluckthun,(1999) FEBS Lett. 459:166-172; Mathys, et al., (1999) Gene 231:1-13;Mills, et al., (1998) Proc. Natl. Acad. Sci. USA 95:3543-3548; Muir, etal., (1998) Proc. Natl. Acad. Sci. USA 95:6705-6710; Otomo, et al.,(1999) Biochemistry 38:16040-16044; Otomo, et al., (1999) J. Biolmol.NMR 14:105-114; Scott, et al., (1999) Proc. Natl. Acad. Sci. USA96:13638-13643; Severinov and Muir, (1998) J. Biol. Chem.273:16205-16209; Shingledecker, et al., (1998) Gene 207:187-195;Southworth, et al., (1998) EMBO J. 17:918-926; Southworth, et al.,(1999) Biotechniques 27:110-120; Wood, et al., (1999) Nat. Biotechnol.17:889-892; Wu, et al., (1998a) Proc. Natl. Acad. Sci. USA 95:9226-9231;Wu, et al., (1998b) Biochim Biophys Acta 1387:422-432; Xu, et al.,(1999) Proc. Natl. Acad. Sci. USA 96:388-393; Yamazaki, et al., (1998)J. Am. Chem. Soc., 120:5591-5592). For the application of inteins inplant transgenes, see, Yang, et al., (Transgene Res 15:583-593 (2006))and Evans, et al., (Annu. Rev. Plant Biol. 56:375-392 (2005)).

In another embodiment, the IPD113 polypeptide may be encoded by twoseparate genes where the intein of the precursor protein comes from thetwo genes, referred to as a split-intein, and the two portions of theprecursor are joined by a peptide bond formation. This peptide bondformation is accomplished by intein-mediated trans-splicing. For thispurpose, a first and a second expression cassette comprising the twoseparate genes further code for inteins capable of mediating proteintrans-splicing. By trans-splicing, the proteins and polypeptides encodedby the first and second fragments may be linked by peptide bondformation. Trans-splicing inteins may be selected from the nucleolar andorganellar genomes of different organisms including eukaryotes,archaebacteria and eubacteria. Inteins that may be used for are listedat neb.com/neb/inteins.html, which can be accessed on the world-wide webusing the “www” prefix). The nucleotide sequence coding for an inteinmay be split into a 5′ and a 3′ part that code for the 5′ and the 3′part of the intein, respectively. Sequence portions not necessary forintein splicing (e.g. homing endonuclease domain) may be deleted. Theintein coding sequence is split such that the 5′ and the 3′ parts arecapable of trans-splicing. For selecting a suitable splitting site ofthe intein coding sequence, the considerations published by Southworth,et al., (1998) EMBO J. 17:918-926 may be followed. In constructing thefirst and the second expression cassette, the 5′ intein coding sequenceis linked to the 3′ end of the first fragment coding for the N-terminalpart of the IPD113 polypeptide and the 3′ intein coding sequence islinked to the 5′ end of the second fragment coding for the C-terminalpart of the IPD113 polypeptide.

In general, the trans-splicing partners can be designed using any splitintein, including any naturally-occurring or artificially-split splitintein. Several naturally-occurring split inteins are known, forexample: the split intein of the DnaE gene of Synechocystis sp. PCC6803(see, Wu, et al., (1998) Proc Natl Acad Sci USA. 95(16):9226-31 andEvans, et al., (2000) J Biol Chem. 275(13):9091-4 and of the DnaE genefrom Nostoc punctiforme (see, Iwai, et al., (2006) FEBS Lett.580(7):1853-8). Non-split inteins have been artificially split in thelaboratory to create new split inteins, for example: the artificiallysplit Ssp DnaB intein (see, Wu, et al., (1998) Biochim Biophys Acta.1387:422-32) and split Sce VMA intein (see, Brenzel, et al., (2006)Biochemistry. 45(6):1571-8) and an artificially split fungal mini-intein(see, Elleuche, et al., (2007) Biochem Biophys Res Commun.355(3):830-4). There are also intein databases available that catalogueknown inteins (see for example the online-database available at:bioinformatics.weizmann.ac.il/{tilde over( )}pietro/inteins/Inteinstable.html, which can be accessed on theworld-wide web using the “www” prefix).

Naturally-occurring non-split inteins may have endonuclease or otherenzymatic activities that can typically be removed when designing anartificially-split split intein. Such mini-inteins or minimized splitinteins are typically less than 200 amino acid residues long (see, Wu,et al., (1998) Biochim Biophys Acta. 1387:422-32). Suitable splitinteins may have other purification enabling polypeptide elements addedto their structure, if such elements do not inhibit the splicing of thesplit intein or are added in a manner that allows them to be removedprior to splicing. Protein splicing has been reported using proteinsthat comprise bacterial intein-like (BIL) domains (see, Amitai, et al.,(2003) Mol Microbiol. 47:61-73) and hedgehog (Hog) auto-processingdomains (the latter is combined with inteins when referred to as theHog/intein superfamily or HINT family (see, Dassa, et al., (2004) J BiolChem. 279:32001-7) and domains such as these may also be used to prepareartificially-split inteins. In particular, non-splicing members of suchfamilies may be modified by molecular biology methodologies to introduceor restore splicing activity in such related species. Recent studiesdemonstrate that splicing can be observed when a N-terminal split inteincomponent can react with a C-terminal split intein component not foundin nature to be its “partner”; for example, splicing has been observedutilizing partners that have as little as 30 to 50% homology with the“natural” splicing partner (see, Dassa, et al., (2007) Biochemistry.46(1):322-30). Other such mixtures of disparate split intein partnershave been shown to be unreactive one with another (see, Brenzel, et al.,(2006) Biochemistry. 45(6):1571-8). However, it is within the ability ofa person skilled in the relevant art to determine whether a pair ofpolypeptides can associate with each other to provide a functionalintein, using routine methods and without the exercise of inventiveskill.

In some embodiments, the IPD113 polypeptide is a circular permutedvariant. In certain embodiments, the IPD113 polypeptide is a circularpermuted variant of the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ IDNO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ IDNO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22,SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO:27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ IDNO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41,SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO:46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ IDNO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60,SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO:65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ IDNO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID

NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82,SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO:87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ IDNO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ IDNO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110,SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ IDNO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119,SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ IDNO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254,SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ IDNO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263,SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ IDNO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272,SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ IDNO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281,SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ IDNO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319,SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ IDNO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426,SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ IDNO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435,SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ IDNO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448,SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ IDNO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466,SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ IDNO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479,SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ IDNO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488,SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ IDNO: 494 or SEQ ID NO: 495, or variant thereof having an amino acidsubstitution, deletion, addition or combinations thereof. Thedevelopment of recombinant DNA methods has made it possible to study theeffects of sequence transposition on protein folding, structure andfunction. The approach used in creating new sequences resembles that ofnaturally occurring pairs of proteins that are related by linearreorganization of their amino acid sequences (Cunningham, et al., (1979)Proc. Natl. Acad. Sci. U.S.A. 76:3218-3222; Teather and Erfle, (1990) J.Bacteriol. 172:3837-3841; Schimming, et al., (1992) Eur. J. Biochem.204:13-19; Yamiuchi and Minamikawa, (1991) FEBS Lett. 260:127-130;MacGregor, et al., (1996) FEBS Lett. 378:263-266). The first in vitroapplication of this type of rearrangement to proteins was described byGoldenberg and Creighton (J. Mol. Biol. 165:407-413, 1983). In creatinga circular permuted variant, a new N-terminus is selected at an internalsite (breakpoint) of the original sequence, the new sequence having thesame order of amino acids as the original from the breakpoint until itreaches an amino acid that is at or near the original C-terminus. Atthis point the new sequence is joined, either directly or through anadditional portion of sequence (linker), to an amino acid that is at ornear the original N-terminus and the new sequence continues with thesame sequence as the original until it reaches a point that is at ornear the amino acid that was N-terminal to the breakpoint site of theoriginal sequence, this residue forming the new C-terminus of the chain.The length of the amino acid sequence of the linker can be selectedempirically or with guidance from structural information or by using acombination of the two approaches. When no structural information isavailable, a small series of linkers can be prepared for testing using adesign whose length is varied to span a range from 0 to 50 Å and whosesequence is chosen to be consistent with surface exposure(hydrophilicity, Hopp and Woods, (1983) Mol. Immunol. 20:483-489; Kyteand Doolittle, (1982) J. Mol. Biol. 157:105-132; solvent exposed surfacearea, Lee and Richards, (1971) J. Mol. Biol. 55:379-400) and the abilityto adopt the necessary conformation without deranging the configurationof the pesticidal polypeptide (conformationally flexible; Karplus andSchulz, (1985) Naturwissenschaften 72:212-213). Assuming an average oftranslation of 2.0 to 3.8 A per residue, this would mean the length totest would be between 0 to 30 residues, with 0 to 15 residues being thepreferred range. Exemplary of such an empirical series would be toconstruct linkers using a cassette sequence such as Gly-Gly-Gly-Serrepeated n times, where n is 1, 2, 3 or 4. Those skilled in the art willrecognize that there are many such sequences that vary in length orcomposition that can serve as linkers with the primary considerationbeing that they be neither excessively long nor short (cf., Sandhu,(1992) Critical Rev. Biotech. 12:437-462); if they are too long, entropyeffects will likely destabilize the three-dimensional fold, and may alsomake folding kinetically impractical, and if they are too short, theywill likely destabilize the molecule because of torsional or stericstrain. Those skilled in the analysis of protein structural informationwill recognize that using the distance between the chain ends, definedas the distance between the c-alpha carbons, can be used to define thelength of the sequence to be used or at least to limit the number ofpossibilities that must be tested in an empirical selection of linkers.They will also recognize that it is sometimes the case that thepositions of the ends of the polypeptide chain are ill-defined instructural models derived from x-ray diffraction or nuclear magneticresonance spectroscopy data, and that when true, this situation willtherefore need to be considered to properly estimate the length of thelinker required. From those residues, whose positions are well definedare selected two residues that are close in sequence to the chain ends,and the distance between their c-alpha carbons is used to calculate anapproximate length for a linker between them. Using the calculatedlength as a guide, linkers with a range of number of residues(calculated using 2 to 3.8 Å per residue) are then selected. Theselinkers may be composed of the original sequence, shortened orlengthened as necessary, and when lengthened the additional residues maybe chosen to be flexible and hydrophilic as described above; oroptionally the original sequence may be substituted for using a seriesof linkers, one example being the Gly-Gly-Gly-Ser cassette approachmentioned above; or optionally a combination of the original sequenceand new sequence having the appropriate total length may be used.Sequences of pesticidal polypeptides capable of folding to biologicallyactive states can be prepared by appropriate selection of the beginning(amino terminus) and ending (carboxyl terminus) positions from withinthe original polypeptide chain while using the linker sequence asdescribed above. Amino and carboxyl termini are selected from within acommon stretch of sequence, referred to as a breakpoint region, usingthe guidelines described below. A novel amino acid sequence is thusgenerated by selecting amino and carboxyl termini from within the samebreakpoint region. In many cases the selection of the new termini willbe such that the original position of the carboxyl terminus immediatelypreceded that of the amino terminus. However, those skilled in the artwill recognize that selections of termini anywhere within the region mayfunction, and that these will effectively lead to either deletions oradditions to the amino or carboxyl portions of the new sequence. It is acentral tenet of molecular biology that the primary amino acid sequenceof a protein dictates folding to the three-dimensional structurenecessary for expression of its biological function. Methods are knownto those skilled in the art to obtain and interpret three-dimensionalstructural information using x-ray diffraction of single proteinCrystals or nuclear magnetic resonance spectroscopy of proteinsolutions. Examples of structural information that are relevant to theidentification of breakpoint regions include the location and type ofprotein secondary structure (alpha and 3-10 helices, parallel andanti-parallel beta sheets, chain reversals and turns, and loops; Kabschand Sander, (1983) Biopolymers 22:2577-2637; the degree of solventexposure of amino acid residues, the extent and type of interactions ofresidues with one another (Chothia, (1984) Ann. Rev. Biochem.53:537-572) and the static and dynamic distribution of conformationsalong the polypeptide chain (Alber and Mathews, (1987) Methods Enzymol.154:511-533). In some cases, additional information is known aboutsolvent exposure of residues; one example is a site ofpost-translational attachment of carbohydrate which is necessarily onthe surface of the protein. When experimental structural information isnot available or is not feasible to obtain, methods are also availableto analyze the primary amino acid sequence to make predictions ofprotein tertiary and secondary structure, solvent accessibility and theoccurrence of turns and loops. Biochemical methods are also sometimesapplicable for empirically determining surface exposure when directstructural methods are not feasible; for example, using theidentification of sites of chain scission following limited proteolysisto infer surface exposure (Gentile and Salvatore, (1993) Eur. J.Biochem. 218:603-621). Thus, using either the experimentally derivedstructural information or predictive methods (e.g., Srinivisan and Rose,(1995) Proteins: Struct., Funct. & Genetics 22:81-99) the parental aminoacid sequence is inspected to classify regions according to whether theyare integral to the maintenance of secondary and tertiary structure. Theoccurrence of sequences within regions that are known to be involved inperiodic secondary structure (alpha and 3-10 helices, parallel andanti-parallel beta sheets) are regions that should be avoided.Similarly, regions of amino acid sequence that are observed or predictedto have a low degree of solvent exposure are more likely to be part ofthe so-called hydrophobic core of the protein and should also be avoidedfor selection of amino and carboxyl termini. In contrast, those regionsthat are known or predicted to be in surface turns or loops, andespecially those regions that are known not to be required forbiological activity, are the preferred sites for location of theextremes of the polypeptide chain. Continuous stretches of amino acidsequence that are preferred based on the above criteria are referred toas a breakpoint region. Polynucleotides encoding circular permutedIPD113 polypeptides with new N-terminus/C-terminus which contain alinker region separating the original C-terminus and N-terminus can bemade essentially following the method described in Mullins, et al.,(1994) J. Am. Chem. Soc. 116:5529-5533. Multiple steps of polymerasechain reaction (PCR) amplifications are used to rearrange the DNAsequence encoding the primary amino acid sequence of the protein.Polynucleotides encoding circular permuted IPD113 polypeptides with newN-terminus/C-terminus which contain a linker region separating theoriginal C-terminus and N-terminus can be made based on thetandem-duplication method described in Horlick, et al., (1992) ProteinEng. 5:427-431. Polymerase chain reaction (PCR) amplification of the newN-terminus/C-terminus genes is performed using a tandemly duplicatedtemplate DNA.

In another embodiment, fusion proteins are provided that include withinits amino acid sequence an amino acid sequence comprising an IPD113polypeptide or chimeric IPD113 polypeptide of the disclosure.Polynucleotides encoding an IPD113 polypeptide may be fused to signalsequences which will direct the localization of the IPD113 polypeptideto particular compartments of a prokaryotic or eukaryotic cell and/ordirect the secretion of the IPD113 polypeptide of the embodiments from aprokaryotic or eukaryotic cell. For example, in E. coli, one may wish todirect the expression of the protein to the periplasmic space. Examplesof signal sequences or proteins (or fragments thereof) to which theIPD113 polypeptide may be fused to direct the expression of thepolypeptide to the periplasmic space of bacteria include, but are notlimited to, the pelB signal sequence, the maltose binding protein (MBP)signal sequence, MBP, the ompA signal sequence, the signal sequence ofthe periplasmic E. coli heat-labile enterotoxin B-subunit and the signalsequence of alkaline phosphatase. Several vectors are commerciallyavailable for the construction of fusion proteins which will direct thelocalization of a protein, such as the pMAL series of vectors(particularly the pMAL-p series) available from New England Biolabs. Ina specific embodiment, the IPD113 polypeptide may be fused to the pelBpectate lyase signal sequence to increase the efficiency of expressionand purification of such polypeptides in Gram-negative bacteria (see,U.S. Pat. Nos. 5,576,195 and 5,846,818). The IPD113 polypeptides of theembodiments may be fused to a plant plastid transit peptide or apoplasttransit peptides such as rice or barley alpha-amylase secretion signal.The plastid transit peptide is generally fused N-terminal to thepolypeptide to be targeted (e.g., the fusion partner). In oneembodiment, the fusion protein consists essentially of the plastidtransit peptide and the IPD113 polypeptide to be targeted. In anotherembodiment, the fusion protein comprises the plastid transit peptide andthe polypeptide to be targeted. In such embodiments, the plastid transitpeptide is preferably at the N-terminus of the fusion protein. However,additional amino acid residues may be N-terminal to the plastid transitpeptide if the fusion protein is at least partially targeted to aplastid. In a specific embodiment, the plastid transit peptide is in theN-terminal half, N-terminal third or N-terminal quarter of the fusionprotein. Most or all of the plastid transit peptide is generally cleavedfrom the fusion protein upon insertion into the plastid. The position ofcleavage may vary slightly between plant species, at different plantdevelopmental stages, because of specific intercellular conditions orthe combination of transit peptide/fusion partner used. In oneembodiment, the plastid transit peptide cleavage is homogenous such thatthe cleavage site is identical in a population of fusion proteins. Inanother embodiment, the plastid transit peptide is not homogenous, suchthat the cleavage site varies by 1-10 amino acids in a population offusion proteins. The plastid transit peptide can be recombinantly fusedto a second protein in one of several ways. For example, a restrictionendonuclease recognition site can be introduced into the nucleotidesequence of the transit peptide at a position corresponding to itsC-terminal end and the same or a compatible site can be engineered intothe nucleotide sequence of the protein to be targeted at its N-terminalend. Care must be taken in designing these sites to ensure that thecoding sequences of the transit peptide and the second protein are kept“in frame” to allow the synthesis of the desired fusion protein. In somecases, it may be preferable to remove the initiator methionine of thesecond protein when the new restriction site is introduced. Theintroduction of restriction endonuclease recognition sites on bothparent molecules and their subsequent joining through recombinant DNAtechniques may result in the addition of one or more extra amino acidsbetween the transit peptide and the second protein. This generally doesnot affect targeting activity if the transit peptide cleavage siteremains accessible and the function of the second protein is not alteredby the addition of these extra amino acids at its N-terminus.Alternatively, one skilled in the art can create a precise cleavage sitebetween the transit peptide and the second protein (with or without itsinitiator methionine) using gene synthesis (Stemmer, et al., (1995) Gene164:49-53) or similar methods. In addition, the transit peptide fusioncan intentionally include amino acids downstream of the cleavage site.The amino acids at the N-terminus of the mature protein can affect theability of the transit peptide to target proteins to plastids and/or theefficiency of cleavage following protein import.

This may be dependent on the protein to be targeted. See, e.g., Comai,et al., (1988) J. Biol. Chem. 263(29):15104-9. In some embodiments, theIPD113 polypeptide is fused to a heterologous signal peptide orheterologous transit peptide.

In some embodiments, fusion proteins are provide comprising an IPD113polypeptide or chimeric IPD113 polypeptide of the disclosure representedby a formula selected from the group consisting of:

R¹-L-R², R²-L-R¹, R¹—R² or R²—R¹

wherein R¹ is an IPD113 polypeptide or chimeric IPD113 polypeptide ofthe disclosure and R² is a protein of interest. In some embodiments, R¹and R² are an IPD113 polypeptide or chimeric IPD113 polypeptide of thedisclosure. The R¹ polypeptide is fused either directly or through alinker (L) segment to the R² polypeptide. The term “directly” definesfusions in which the polypeptides are joined without a peptide linker.Thus “L” represents a chemical bound or polypeptide segment to whichboth R¹ and R² are fused in frame, most commonly L is a linear peptideto which R¹ and R² are bound by amide bonds linking the carboxy terminusof R¹ to the amino terminus of L and carboxy terminus of L to the aminoterminus of R². By “fused in frame” is meant that there is notranslation termination or disruption between the reading frames of R¹and R². The linking group (L) is generally a polypeptide of between 1and 500 amino acids in length. The linkers joining the two molecules arepreferably designed to (1) allow the two molecules to fold and actindependently of each other, (2) not have a propensity for developing anordered secondary structure which could interfere with the functionaldomains of the two proteins, (3) have minimal hydrophobic or chargedcharacteristic which could interact with the functional protein domainsand (4) provide steric separation of R¹ and R² such that R¹ and R² couldinteract simultaneously with their corresponding receptors on a singlecell. Typically surface amino acids in flexible protein regions includeGly, Asn and Ser. Virtually any permutation of amino acid sequencescontaining Gly, Asn and Ser would be expected to satisfy the abovecriteria for a linker sequence. Other neutral amino acids, such as Thrand Ala, may also be used in the linker sequence. Additional amino acidsmay also be included in the linkers due to the addition of uniquerestriction sites in the linker sequence to facilitate construction ofthe fusions.

In some embodiments, the linkers comprise sequences selected from thegroup of formulas: (Gly₃Ser)_(n), (Gly4Ser)_(n), (Gly₅Ser)_(n),(GlynSer)_(n) or (AlaGlySer)_(n) where n is an integer. One example of ahighly-flexible linker is the (GlySer)-rich spacer region present withinthe pill protein of the filamentous bacteriophages, e.g. bacteriophagesM13 or fd (Schaller, et al., 1975). This region provides a long,flexible spacer region between two domains of the pill surface protein.Also included are linkers in which an endopeptidase recognition sequenceis included. Such a cleavage site may be valuable to separate theindividual components of the fusion to determine if they are properlyfolded and active in vitro. Examples of various endopeptidases include,but are not limited to, Plasmin, Enterokinase, Kallikerin, Urokinase,Tissue Plasminogen activator, clostripain, Chymosin, Collagenase,Russell's Viper Venom Protease, Postproline cleavage enzyme, V8protease, Thrombin and factor Xa. In some embodiments, the linkercomprises the amino acids EEKKN (SEQ ID NO: 334) from the multi-geneexpression vehicle (MGEV), which is cleaved by vacuolar proteases asdisclosed in US Patent Application Publication Number US 2007/0277263.In other embodiments, peptide linker segments from the hinge region ofheavy chain immunoglobulins IgG, IgA, IgM, IgD or IgE provide an angularrelationship between the attached polypeptides. Especially useful arethose hinge regions where the cysteines are replaced with serines.Linkers of the present disclosure include sequences derived from murineIgG gamma 2b hinge region in which the cysteines have been changed toserines. The fusion proteins are not limited by the form, size or numberof linker sequences employed and the only requirement of the linker isthat functionally it does not interfere adversely with the folding andfunction of the individual molecules of the fusion.

Nucleic Acid Molecules, and Variants and Fragments Thereof

Isolated or recombinant nucleic acid molecules comprising nucleic acidsequences encoding IPD113 polypeptides or biologically active portionsthereof, as well as nucleic acid molecules sufficient for use ashybridization probes to identify nucleic acid molecules encodingproteins with regions of sequence homology are provided. As used herein,the term “nucleic acid molecule” refers to DNA molecules (e.g.,recombinant DNA, cDNA, genomic DNA, plastid DNA, mitochondrial DNA) andRNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated usingnucleotide analogs. The nucleic acid molecule can be single-stranded ordouble-stranded, but preferably is double-stranded DNA.

An “isolated” nucleic acid molecule (or DNA) is used herein to refer toa nucleic acid sequence (or DNA) that is no longer in its naturalenvironment, for example in vitro. A “recombinant” nucleic acid molecule(or DNA) is used herein to refer to a nucleic acid sequence (or DNA)that is in a recombinant bacterial or plant host cell. In someembodiments, an “isolated” or “recombinant” nucleic acid is free ofsequences (preferably protein encoding sequences) that naturally flankthe nucleic acid (i.e., sequences located at the 5′ and 3′ ends of thenucleic acid) in the genomic DNA of the organism from which the nucleicacid is derived. For purposes of the disclosure, “isolated” or“recombinant” when used to refer to nucleic acid molecules excludesisolated chromosomes. For example, in various embodiments, therecombinant nucleic acid molecules encoding IPD113 polypeptides cancontain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kbof nucleic acid sequences that naturally flank the nucleic acid moleculein genomic DNA of the cell from which the nucleic acid is derived.

In some embodiments, an isolated nucleic acid molecule encoding IPD113polypeptides has one or more change in the nucleic acid sequencecompared to the native or genomic nucleic acid sequence. In someembodiments, the change in the native or genomic nucleic acid sequenceincludes but is not limited to: changes in the nucleic acid sequence dueto the degeneracy of the genetic code; changes in the nucleic acidsequence due to the amino acid substitution, insertion, deletion and/oraddition compared to the native or genomic sequence; removal of one ormore intron; deletion of one or more upstream or downstream regulatoryregions; and deletion of the 5′ and/or 3′ untranslated region associatedwith the genomic nucleic acid sequence. In some embodiments, the nucleicacid molecule encoding an IPD113 polypeptide is a non-genomic sequence.

A variety of polynucleotides that encode IPD113 polypeptides or relatedproteins are contemplated. Such polynucleotides are useful forproduction of IPD113 polypeptides in host cells when operably linked toa suitable promoter, transcription termination and/or polyadenylationsequences. Such polynucleotides are also useful as probes for isolatinghomologous or substantially homologous polynucleotides that encodeIPD113 polypeptides or related proteins.

Methods for Engineering IPD113 Polypeptides

Methods for engineering IPD113 polypeptides are also encompassed by thedisclosure. In some embodiments, the method for engineering IPD113polypeptides uses rational protein design based on a secondary, tertiaryor quaternary structure model of the IPD113 polypeptide. In-silicomodeling tools can be used in the methods of the disclosure. In someembodiments, the rational protein design uses an in-silico modeling toolselected from, but not limited to, PyMOL (PyMOL Molecular GraphicsSystem, Version 1.7.4 Schrödinger, LLC.), Maestro©, BioLuminate (Zhu,K.; et al., Proteins, 2014, 82(8), 1646-1655; Salam, N. K et al.,Protein Eng. Des. Sel., 2014, 27(10), 365-74; Beard, H. et al. PLoS ONE,2013, 8(12), e82849), MOE© (Molecular Operating Environment (MOE),2013.08; Chemical Computing Group Inc., 1010 Sherbooke St. West, Suite#910, Montreal, QC, Canada, H3A 2R7, 2015), Jmol, and Discovery Studio©(Accelrys Software Inc. Discovery Studio Modeling Environment, Release3.5.0, San Diego: Accelrys Software Inc. 2013). In some embodiments, themodeling uses Discovery Studio© software. In some embodiments, themethod the structural coordinates can be determined by homologymodeling. In some embodiments, the method the structural coordinates canbe determined by X-ray crystallography or solution NMR.

In some embodiments, the IPD113 polypeptide is engineered by the methodof the disclosure to have a modified physical property compared to thenative IPD113 polypeptide. In some embodiments, the modified physicalproperties include, but are not limited to net surface charge and chargedistribution on the protein surface, net hydrophobicity and hydrophobicresidue distribution on the protein surface, surface charge density,surface hydrophobicity density, total count of surface ionizable groups,and protein size. In some embodiments, the modified physicalin-properties include, but are not limited to solubility, folding,stability, protease stability, digestibility, planta expression,insecticidal potency, spectrum of insecticidal activity, ion channelactivity of protomer pore, and receptor binding. In some embodiments,the modified physical property is improved protease stability, improvedin-planta expression, improved solubility, improved potency, improvedion-channel activity of protomer pore, and/or improved receptor binding.

Using the methods of the disclosure, proteolytically-sensitive sites canbe identified and may be modified or utilized to produce more stable ormore biologically active IPD113 polypeptides.

Using methods of the disclosure, sites involved in receptor bindingand/or pore formation can be identified and may be modified to createIPD113 polypeptides having enhanced insecticidal activity; enhancedability to form channels; and reduced size.

Using methods of the disclosure, occupation of a site by a watermolecule can be identified and can be modified to create IPD113molecules having modified flexibility in a region or increasing thenumber of hydrophobic residues along that surface, which may be involvedin receptor binding and/or pore formation.

Using methods of the disclosure, hydrogen bonding in a region can beidentified and the amino acids may be substituted to modify the numberof hydrogen bonds, including salt bridges, to create IPD113 polypeptideshaving a modified hydrophobic interaction surface facilitating pre-poreand pore formation and/or modified insecticidal activity.

Using methods of the disclosure, loop regions can be identified and maybe modified to create IPD113 polypeptides having modified channel orpore formation, folding, and/or receptor binding.

Using methods of the disclosure, complex electrostatic surfaces andhydrophobic or hydrophilic interactions can be identified and modifiedto create IPD113 polypeptides having modified receptor interaction

Using methods of the disclosure, metal binding sites can be identifiedand modified to create IPD113 polypeptides having modified ion channelor pore activity.

Using methods of the disclosure, amino acids that may be buried orotherwise removed from the surface of the protein that hold in place thethree-dimensional structure can be identified and modified to createIPD113 polypeptides having modified stability or flexibility.

Using methods of the disclosure, non-specific binding sites to otherbiomolecules can be identified and modified to create IPD113polypeptides having modified receptor binding to the specific receptorand enhanced toxicity.

Appling various computational tools coupled with the understanding ofsaturated mutagenesis, and the structural/functional relationship forIPD113 polypeptides as disclosed herein, one skilled in the art canidentify and modify various physical properties of IPD113 polypeptidesfor the better overall performance as an insecticidal protein againstthe desired targets. Combinatory mutagenesis at various regions canenhance specificity to the current active targets and potentially canalso change activity spectrum against different targets. Such targetedcombinatorial mutagenesis can be achieved with incorporation ofmutagenic oligo nucleotides or generated by gene synthesis or thecombination of both approaches. Mutagenesis on defined loop regions canalso enhance physical properties of IPD113 polypeptides such asincreasing protein stability by reducing protease degradation abilityand increasing thermostability etc. In additional, combinatorialmutagenesis can be applied to the amino acid residues involved inhydrophobic interface surface. Enhancement of hydrophobic interfacesurface can potentially increase insecticidal activity, thermostabilityand other physical properties. Additional improvements can also beachieved through mutagenesis of other part of the molecule such asvarious beta-sheets and alpha helices to increase stability andactivity.

Polynucleotides Encoding IPD113 Polypeptides

One source of polynucleotides that encode IPD113 polypeptides or relatedproteins is a fern or other primitive plant species selected from, butnot limited to, limited to Pteris, Polypodium, Nephrolepis, Colysis,Tectaria, Davallia, Polystichum, Adiantum, Asplenium, Blechnum,Lygodium, Ophioglossum, Pyrrosia, Doryopteris, Dryopteris, Pellaea,Gymnocarpium, Cheilanthes, Pteridium, Christella, Lastreopsis,Campyloneurum, Hemionitis, Selliguea, and Arachniodes species, whichcontains an IPD113 polynucleotide of SEQ ID NO: 127, SEQ ID NO: 128, SEQID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO:133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO:142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO:151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO:160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO:169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO:178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 182, SEQID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO:187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO:196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO:205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO:214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 217, SEQ ID NO: 218, SEQID NO: 219, SEQ ID NO: 220, SEQ ID NO: 221, SEQ ID NO: 222, SEQ ID NO:223, SEQ ID NO: 224, SEQ ID NO: 225, SEQ ID NO: 226, SEQ ID NO: 227, SEQID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO:232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, SEQ ID NO: 240, SEQ ID NO:241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ ID NO: 244, SEQ ID NO: 245, SEQID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO:250, SEQ ID NO: 251, SEQ ID NO: 252, SEQ ID NO: 282, SEQ ID NO: 283, SEQID NO: 284, SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO:288, SEQ ID NO: 289, SEQ ID NO: 290, SEQ ID NO: 291, SEQ ID NO: 292, SEQID NO: 293, SEQ ID NO: 294, SEQ ID NO: 295, SEQ ID NO: 296, SEQ ID NO:297, SEQ ID NO: 298, SEQ ID NO: 299, SEQ ID NO: 300, SEQ ID NO: 301, SEQID NO: 302, SEQ ID NO: 303, SEQ ID NO: 304, SEQ ID NO: 305, SEQ ID NO:306, SEQ ID NO: 307, SEQ ID NO: 308, SEQ ID NO: 309. SEQ ID NO: 310, SEQID NO: 321, SEQ ID NO: 322, SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO:325, SEQ ID NO: 326, SEQ ID NO: 327, SEQ ID NO: 328, SEQ ID NO: 329, SEQID NO: 330, SEQ ID NO: 335, SEQ ID NO: 338, SEQ ID NO: 339, SEQ ID NO:341, SEQ ID NO: 342, SEQ ID NO: 343, SEQ ID NO: 344, SEQ ID NO: 345, SEQID NO: 346, SEQ ID NO: 347, SEQ ID NO: 348, SEQ ID NO: 349, SEQ ID NO:350, SEQ ID NO: 351, SEQ ID NO: 352, SEQ ID NO: 353, SEQ ID NO: 354, SEQID NO: 355, SEQ ID NO: 357, SEQ ID NO: 358, SEQ ID NO: 359, SEQ ID NO:360, SEQ ID NO: 362, SEQ ID NO: 364, SEQ ID NO: 366, SEQ ID NO: 367, SEQID NO: 369, SEQ ID NO: 370, SEQ ID NO: 371, SEQ ID NO: 372, SEQ ID NO:377, SEQ ID NO: 379, SEQ ID NO: 380, SEQ ID NO: 382, SEQ ID NO: 385, SEQID NO: 388, SEQ ID NO: 389, SEQ ID NO: 391, SEQ ID NO: 392, SEQ ID NO:393, SEQ ID NO: 394, SEQ ID NO: 396, SEQ ID NO: 397, SEQ ID NO: 398, SEQID NO: 399, SEQ ID NO: 400, SEQ ID NO: 401, SEQ ID NO: 402, SEQ ID NO:403, SEQ ID NO: 404, SEQ ID NO: 405, SEQ ID NO: 406, SEQ ID NO: 407, SEQID NO: 408, SEQ ID NO: 409, SEQ ID NO: 410, SEQ ID NO: 411, SEQ ID NO:413, and SEQ ID NO: 414 encoding an IPD113 polypeptide of SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6,SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11,SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ IDNO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30,SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO:35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ IDNO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO:54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ IDNO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68,SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO:73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ IDNO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO:92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ IDNO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ IDNO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110,SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ IDNO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119,SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ IDNO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254,SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ IDNO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263,SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ IDNO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272,SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ IDNO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281,SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ IDNO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319,SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ IDNO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426,SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ IDNO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435,SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ IDNO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448,SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ IDNO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466,SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ IDNO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479,SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ IDNO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488,SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ IDNO: 494, and SEQ ID NO: 495 respectively.

The polynucleotides of SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129,SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ IDNO: 134, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138,SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, SEQ IDNO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147,SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ IDNO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156,SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ IDNO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165,SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ IDNO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174,SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ IDNO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183,SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ IDNO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192,SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ IDNO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201,SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ IDNO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210,SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ IDNO: 215, SEQ ID NO: 216, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219,SEQ ID NO: 220, SEQ ID NO: 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ IDNO: 224, SEQ ID NO: 225, SEQ ID NO: 226, SEQ ID NO: 227, SEQ ID NO: 228,SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ IDNO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237,SEQ ID NO: 238, SEQ ID NO: 239, SEQ ID NO: 240, SEQ ID NO: 241, SEQ IDNO: 242, SEQ ID NO: 243, SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 246,SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 250, SEQ IDNO: 251, SEQ ID NO: 252, SEQ ID NO: 282, SEQ ID NO: 283, SEQ ID NO: 284,SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO: 288, SEQ IDNO: 289, SEQ ID NO: 290, SEQ ID NO: 291, SEQ ID NO: 292, SEQ ID NO: 293,SEQ ID NO: 294, SEQ ID NO: 295, SEQ ID NO: 296, SEQ ID NO: 297, SEQ IDNO: 298, SEQ ID NO: 299, SEQ ID NO: 300, SEQ ID NO: 301, SEQ ID NO: 302,SEQ ID NO: 303, SEQ ID NO: 304, SEQ ID NO: 305, SEQ ID NO: 306, SEQ IDNO: 307, SEQ ID NO: 308, SEQ ID NO: 309. SEQ ID NO: 310, SEQ ID NO: 321,SEQ ID NO: 322, SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 325, SEQ IDNO: 326, SEQ ID NO: 327, SEQ ID NO: 328, SEQ ID NO: 329, SEQ ID NO: 330,SEQ ID NO: 335, SEQ ID NO: 338, SEQ ID NO: 339, SEQ ID NO: 341, SEQ IDNO: 342, SEQ ID NO: 343, SEQ ID NO: 344, SEQ ID NO: 345, SEQ ID NO: 346,SEQ ID NO: 347, SEQ ID NO: 348, SEQ ID NO: 349, SEQ ID NO: 350, SEQ IDNO: 351, SEQ ID NO: 352, SEQ ID NO: 353, SEQ ID NO: 354, SEQ ID NO: 355,SEQ ID NO: 357, SEQ ID NO: 358, SEQ ID NO: 359, SEQ ID NO: 360, SEQ IDNO: 362, SEQ ID NO: 364, SEQ ID NO: 366, SEQ ID NO: 367, SEQ ID NO: 369,SEQ ID NO: 370, SEQ ID NO: 371, SEQ ID NO: 372, SEQ ID NO: 377, SEQ IDNO: 379, SEQ ID NO: 380, SEQ ID NO: 382, SEQ ID NO: 385, SEQ ID NO: 388,SEQ ID NO: 389, SEQ ID NO: 391, SEQ ID NO: 392, SEQ ID NO: 393, SEQ IDNO: 394, SEQ ID NO: 396, SEQ ID NO: 397, SEQ ID NO: 398, SEQ ID NO: 399,SEQ ID NO: 400, SEQ ID NO: 401, SEQ ID NO: 402, SEQ ID NO: 403, SEQ IDNO: 404, SEQ ID NO: 405, SEQ ID NO: 406, SEQ ID NO: 407, SEQ ID NO: 408,SEQ ID NO: 409, SEQ ID NO: 410, SEQ ID NO: 411, SEQ ID NO: 413, and SEQID NO: 414 can be used to express IPD113 polypeptides in recombinantbacterial hosts that include but are not limited to Agrobacterium,Bacillus, Escherichia, Salmonella, Pseudomonas and Rhizobium bacterialhost cells. The polynucleotides are also useful as probes for isolatinghomologous or substantially homologous polynucleotides that encodeIPD113 polypeptides or related proteins. Such probes can be used toidentify homologous or substantially homologous polynucleotides derivedfrom fern or other primitive plant species selected from, but notlimited to, Pteris, Polypodium, Nephrolepis, Colysis, Tectaria,Davallia, Polystichum, Adiantum, Asplenium, Blechnum, Lygodium,Ophioglossum, Pyrrosia, Doryopteris, Dryopteris, Pellaea, Gymnocarpium,Cheilanthes, Pteridium, Christella, Lastreopsis, Campyloneurum,Hemionitis, Selliguea, and Arachniodes species.

Polynucleotides that encode IPD113 polypeptides can also be synthesizedde novo from an IPD113 polypeptide sequence. The sequence of thepolynucleotide gene can be deduced from an IPD113 polypeptide sequencethrough use of the genetic code. Computer programs such as“BackTranslate” (GCG™ Package, Acclerys, Inc. San Diego, Calif.) can beused to convert a peptide sequence to the corresponding nucleotidesequence encoding the peptide. Examples of IPD113 polypeptide sequencesthat can be used to obtain corresponding nucleotide encoding sequencesinclude, but are not limited to the IPD113 polypeptides of SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6,SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11,SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ IDNO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30,SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO:35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ IDNO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO:54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ IDNO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68,SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO:73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ IDNO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO:92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ IDNO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ IDNO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110,SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ IDNO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119,SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ IDNO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254,SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ IDNO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263,SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ IDNO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272,SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ IDNO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281,SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ IDNO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319,SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ IDNO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426,SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ IDNO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435,SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ IDNO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448,SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ IDNO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466,SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ IDNO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479,SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ IDNO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488,SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ IDNO: 494 or SEQ ID NO: 495. Furthermore, synthetic IPD113 polynucleotidesequences of the disclosure can be designed so that they will beexpressed in plants.

In some embodiments, the nucleic acid molecule encoding an IPD113polypeptide is a polynucleotide having the sequence set forth in SEQ IDNO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131,SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ IDNO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140,SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ IDNO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149,SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ IDNO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158,SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ IDNO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167,SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ IDNO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176,SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ IDNO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185,SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ IDNO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194,SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ IDNO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203,SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ IDNO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212,SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ IDNO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO: 221,SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, SEQ IDNO: 226, SEQ ID NO: 227, SEQ ID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230,SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ IDNO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239,SEQ ID NO: 240, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ IDNO: 244, SEQ ID NO: 245, SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248,SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO: 251, SEQ ID NO: 252, SEQ IDNO: 282, SEQ ID NO: 283, SEQ ID NO: 284, SEQ ID NO: 285, SEQ ID NO: 286,SEQ ID NO: 287, SEQ ID NO: 288, SEQ ID NO: 289, SEQ ID NO: 290, SEQ IDNO: 291, SEQ ID NO: 292, SEQ ID NO: 293, SEQ ID NO: 294, SEQ ID NO: 295,SEQ ID NO: 296, SEQ ID NO: 297, SEQ ID NO: 298, SEQ ID NO: 299, SEQ IDNO: 300, SEQ ID NO: 301, SEQ ID NO: 302, SEQ ID NO: 303, SEQ ID NO: 304,SEQ ID NO: 305, SEQ ID NO: 306, SEQ ID NO: 307, SEQ ID NO: 308, SEQ IDNO: 309. SEQ ID NO: 310, SEQ ID NO: 321, SEQ ID NO: 322, SEQ ID NO: 323,SEQ ID NO: 324, SEQ ID NO: 325, SEQ ID NO: 326, SEQ ID NO: 327, SEQ IDNO: 328, SEQ ID NO: 329, SEQ ID NO: 330, SEQ ID NO: 335, SEQ ID NO: 338,SEQ ID NO: 339, SEQ ID NO: 341, SEQ ID NO: 342, SEQ ID NO: 343, SEQ IDNO: 344, SEQ ID NO: 345, SEQ ID NO: 346, SEQ ID NO: 347, SEQ ID NO: 348,SEQ ID NO: 349, SEQ ID NO: 350, SEQ ID NO: 351, SEQ ID NO: 352, SEQ IDNO: 353, SEQ ID NO: 354, SEQ ID NO: 355, SEQ ID NO: 357, SEQ ID NO: 358,SEQ ID NO: 359, SEQ ID NO: 360, SEQ ID NO: 362, SEQ ID NO: 364, SEQ IDNO: 366, SEQ ID NO: 367, SEQ ID NO: 369, SEQ ID NO: 370, SEQ ID NO: 371,SEQ ID NO: 372, SEQ ID NO: 377, SEQ ID NO: 379, SEQ ID NO: 380, SEQ IDNO: 382, SEQ ID NO: 385, SEQ ID NO: 388, SEQ ID NO: 389, SEQ ID NO: 391,SEQ ID NO: 392, SEQ ID NO: 393, SEQ ID NO: 394, SEQ ID NO: 396, SEQ IDNO: 397, SEQ ID NO: 398, SEQ ID NO: 399, SEQ ID NO: 400, SEQ ID NO: 401,SEQ ID NO: 402, SEQ ID NO: 403, SEQ ID NO: 404, SEQ ID NO: 405, SEQ IDNO: 406, SEQ ID NO: 407, SEQ ID NO: 408, SEQ ID NO: 409, SEQ ID NO: 410,SEQ ID NO: 411, SEQ ID NO: 413 or SEQ ID NO: 414 and variants, fragmentsand complements thereof. Nucleic acid sequences that are complementaryto a nucleic acid sequence of the embodiments or that hybridize to asequence of the embodiments are also encompassed. The nucleic acidsequences can be used in DNA constructs or expression cassettes fortransformation and expression in organisms, including microorganisms andplants. The nucleotide or amino acid sequences may be syntheticsequences that have been designed for expression in an organismincluding, but not limited to, a microorganism or a plant.

“Complement” is used herein to refer to a nucleic acid sequence that issufficiently complementary to a given nucleic acid sequence such that itcan hybridize to the given nucleic acid sequence to thereby form astable duplex. “Polynucleotide sequence variants” is used herein torefer to a nucleic acid sequence that except for the degeneracy of thegenetic code encodes the same polypeptide.

In some embodiments, the nucleic acid molecule encoding the IPD113polypeptide is a non-genomic nucleic acid sequence. As used herein a“non-genomic nucleic acid sequence” or “non-genomic nucleic acidmolecule” or “non-genomic polynucleotide” refers to a nucleic acidmolecule that has one or more change in the nucleic acid sequencecompared to a native or genomic nucleic acid sequence. In someembodiments, the change to a native or genomic nucleic acid moleculeincludes but is not limited to: changes in the nucleic acid sequence dueto the degeneracy of the genetic code; optimization of the nucleic acidsequence for expression in plants; changes in the nucleic acid sequenceto introduce at least one amino acid substitution, insertion, deletionand/or addition compared to the native or genomic sequence; removal ofone or more intron associated with the genomic nucleic acid sequence;insertion of one or more heterologous introns; deletion of one or moreupstream or downstream regulatory regions associated with the genomicnucleic acid sequence; insertion of one or more heterologous upstream ordownstream regulatory regions; deletion of the 5′ and/or 3′ untranslatedregion associated with the genomic nucleic acid sequence; insertion of aheterologous 5′ and/or 3′ untranslated region; and modification of apolyadenylation site. In some embodiments, the non-genomic nucleic acidmolecule is a synthetic nucleic acid sequence.

In some embodiments, the nucleic acid molecule encoding an IPD113polypeptide is a non-genomic polynucleotide having a nucleotide sequencehaving at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%,61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%,75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greateridentity, to the nucleic acid sequence of SEQ ID NO: 127, SEQ ID NO:128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO:137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO:146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO:155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO:164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO:173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO:182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO:191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO:200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO:209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 217, SEQ ID NO:218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO: 221, SEQ ID NO: 222, SEQID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, SEQ ID NO: 226, SEQ ID NO:227, SEQ ID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO:236, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, SEQ ID NO: 240, SEQID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ ID NO: 244, SEQ ID NO:245, SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 249, SEQID NO: 250, SEQ ID NO: 251, SEQ ID NO: 252, SEQ ID NO: 282, SEQ ID NO:283, SEQ ID NO: 284, SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQID NO: 288, SEQ ID NO: 289, SEQ ID NO: 290, SEQ ID NO: 291, SEQ ID NO:292, SEQ ID NO: 293, SEQ ID NO: 294, SEQ ID NO: 295, SEQ ID NO: 296, SEQID NO: 297, SEQ ID NO: 298, SEQ ID NO: 299, SEQ ID NO: 300, SEQ ID NO:301, SEQ ID NO: 302, SEQ ID NO: 303, SEQ ID NO: 304, SEQ ID NO: 305, SEQID NO: 306, SEQ ID NO: 307, SEQ ID NO: 308, SEQ ID NO: 309. SEQ ID NO:310, SEQ ID NO: 321, SEQ ID NO: 322, SEQ ID NO: 323, SEQ ID NO: 324, SEQID NO: 325, SEQ ID NO: 326, SEQ ID NO: 327, SEQ ID NO: 328, SEQ ID NO:329, SEQ ID NO: 330, SEQ ID NO: 335, SEQ ID NO: 338, SEQ ID NO: 339, SEQID NO: 341, SEQ ID NO: 342, SEQ ID NO: 343, SEQ ID NO: 344, SEQ ID NO:345, SEQ ID NO: 346, SEQ ID NO: 347, SEQ ID NO: 348, SEQ ID NO: 349, SEQID NO: 350, SEQ ID NO: 351, SEQ ID NO: 352, SEQ ID NO: 353, SEQ ID NO:354, SEQ ID NO: 355, SEQ ID NO: 357, SEQ ID NO: 358, SEQ ID NO: 359, SEQID NO: 360, SEQ ID NO: 362, SEQ ID NO: 364, SEQ ID NO: 366, SEQ ID NO:367, SEQ ID NO: 369, SEQ ID NO: 370, SEQ ID NO: 371, SEQ ID NO: 372, SEQID NO: 377, SEQ ID NO: 379, SEQ ID NO: 380, SEQ ID NO: 382, SEQ ID NO:385, SEQ ID NO: 388, SEQ ID NO: 389, SEQ ID NO: 391, SEQ ID NO: 392, SEQID NO: 393, SEQ ID NO: 394, SEQ ID NO: 396, SEQ ID NO: 397, SEQ ID NO:398, SEQ ID NO: 399, SEQ ID NO: 400, SEQ ID NO: 401, SEQ ID NO: 402, SEQID NO: 403, SEQ ID NO: 404, SEQ ID NO: 405, SEQ ID NO: 406, SEQ ID NO:407, SEQ ID NO: 408, SEQ ID NO: 409, SEQ ID NO: 410, SEQ ID NO: 411, SEQID NO: 413 or SEQ ID NO: 414, wherein the encoded IPD113 polypeptide hasinsecticidal activity.

In some embodiments, the IPD113 polynucleotide encodes an IPD113polypeptide having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%,55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%,69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or greater sequence identity compared to SEQ ID NO: 1, SEQID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO:420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO:430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO:440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO:453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO:473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO:483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO:492, SEQ ID NO: 494, or SEQ ID NO: 495 and has at least one amino acidsubstitution, deletion, insertion or combination therefore, compared tothe native sequence.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 9, SEQID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14,SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO:19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 24, SEQ ID NO: 27, SEQ ID NO: 30, SEQ ID NO: 35, SEQ ID NO: 36, SEQID NO: 37, SEQ ID NO: 38, SEQ ID NO: 77, SEQ ID NO: 88, SEQ ID NO: 89,SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO:95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 100, SEQ IDNO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105,SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 110, SEQ ID NO: 111, SEQ IDNO: 112, SEQ ID NO: 113 or SEQ ID NO: 114.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 24,SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO:103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQID NO: 39 or SEQ ID NO: 40.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 88,SEQ ID NO: 89, SEQ ID NO: 90 or SEQ ID NO: 91.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 20,SEQ ID NO: 24 or SEQ ID NO: 27.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 13,SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 21 or SEQ ID NO: 22.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 17,SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 38, SEQ ID NO: 77, SEQ ID NO:110, SEQ ID NO: 111 or SEQ ID NO: 112.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater identityacross the entire length of the amino acid sequence of SEQ ID NO: 9, SEQID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 23,SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 97, SEQ ID NO:98, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 113 orSEQ ID NO: 114.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 80%,81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or greater identity across the entire length ofthe amino acid sequence of SEQ ID NO: 10.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprises an amino acid sequence having at least about 80%,81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or greater identity across the entire length ofthe amino acid sequence of SEQ ID NO: 16.

In some embodiments, the nucleic acid molecule encodes an IPD113polypeptide comprising an amino acid sequence of SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO:255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO:264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO:273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO:311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO:320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO:427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO:436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO:450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO:469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO:480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO:489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 orSEQ ID NO: 495, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35 or more amino acid substitutions, deletions and/or insertionscompared to the native amino acid at the corresponding position of SEQID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ IDNO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ IDNO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20,SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ IDNO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39,SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO:44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ IDNO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58,SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO:63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ IDNO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77,SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO:82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ IDNO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96,SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO:101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO:110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO:119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO:254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO:263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO:272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO:281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO:319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO:426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO:435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO:448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO:466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO:479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO:488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQID NO: 494 or SEQ ID NO: 495.

Also provided are nucleic acid molecules that encode transcriptionand/or translation products that are subsequently spliced to ultimatelyproduce functional IPD113 polypeptides.

Splicing can be accomplished in vitro or in vivo, and can involve cis-or trans-splicing. The substrate for splicing can be polynucleotides(e.g., RNA transcripts) or polypeptides. An example of cis-splicing of apolynucleotide is where an intron inserted into a coding sequence isremoved and the two flanking exon regions are spliced to generate anIPD113 polypeptide encoding sequence. An example of trans-splicing wouldbe where a polynucleotide is encrypted by separating the coding sequenceinto two or more fragments that can be separately transcribed and thenspliced to form the full-length pesticidal encoding sequence. The use ofa splicing enhancer sequence, which can be introduced into a construct,can facilitate splicing either in cis or trans-splicing of polypeptides(U.S. Pat. Nos. 6,365,377 and 6,531,316). Thus, in some embodiments, thepolynucleotides do not directly encode a full-length IPD113 polypeptide,but rather encode a fragment or fragments of an IPD113 polypeptide.These polynucleotides can be used to express a functional IPD113polypeptide through a mechanism involving splicing, where splicing canoccur at the level of polynucleotide (e.g., intron/exon) and/orpolypeptide (e.g., intein/extein). This can be useful, for example, incontrolling expression of pesticidal activity, since a functionalpesticidal polypeptide will only be expressed if all required fragmentsare expressed in an environment that permits splicing processes togenerate functional product. In another example, introduction of one ormore insertion sequences into a polynucleotide can facilitaterecombination with a low homology polynucleotide; use of an intron orintein for the insertion sequence facilitates the removal of theintervening sequence, thereby restoring function of the encoded variant.

Nucleic acid molecules that are fragments of these nucleic acidsequences encoding IPD113 polypeptides are also encompassed by theembodiments. “Fragment” as used herein refers to a portion of thenucleic acid sequence encoding an IPD113 polypeptide. A fragment of anucleic acid sequence may encode a biologically active portion of anIPD113 polypeptide or it may be a fragment that can be used as ahybridization probe or PCR primer using methods disclosed below. Nucleicacid molecules that are fragments of a nucleic acid sequence encoding anIPD113 polypeptide comprise at least about 150, 180, 210, 240, 270, 300,330 or 360, contiguous nucleotides or up to the number of nucleotidespresent in a full-length nucleic acid sequence encoding an IPD113polypeptide disclosed herein, depending upon the intended use.“Contiguous nucleotides” is used herein to refer to nucleotide residuesthat are immediately adjacent to one another. Fragments of the nucleicacid sequences of the embodiments will encode protein fragments thatretain the biological activity of the IPD113 polypeptide and, hence,retain insecticidal activity. “Retains insecticidal activity” is usedherein to refer to a polypeptide having at least about 10%, at leastabout 30%, at least about 50%, at least about 70%, 80%, 90%, 95% orhigher of the insecticidal activity of the full-length IPD113polypeptide. In some embodiments, the insecticidal activity is against aLepidopteran species.

In some embodiments, the insecticidal activity is against one or moreinsect pests selected from Soy Bean Looper (SBL) (Pseudoplusiaincludes), Fall Armyworm (FAW) (Spodoptera frupperda), Corn Earworm(CEW) (Helicoverpa zea), Velvet Bean Caterpillar (VBC) (Anticarsiagemmatalis) and European Corn Borer (ECB) (Ostrinia nubialis).

In some embodiments, the IPD113 polypeptide is encoded by a nucleic acidsequence sufficiently homologous to the nucleic acid sequence of SEQ IDNO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131,SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ IDNO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140,SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ IDNO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149,SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ IDNO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158,SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ IDNO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167,SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ IDNO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176,SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ IDNO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185,SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ IDNO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194,SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ IDNO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203,SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ IDNO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212,SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ IDNO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO: 221,SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, SEQ IDNO: 226, SEQ ID NO: 227, SEQ ID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230,SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ IDNO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239,SEQ ID NO: 240, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ IDNO: 244, SEQ ID NO: 245, SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248,SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO: 251, SEQ ID NO: 252, SEQ IDNO: 282, SEQ ID NO: 283, SEQ ID NO: 284, SEQ ID NO: 285, SEQ ID NO: 286,SEQ ID NO: 287, SEQ ID NO: 288, SEQ ID NO: 289, SEQ ID NO: 290, SEQ IDNO: 291, SEQ ID NO: 292, SEQ ID NO: 293, SEQ ID NO: 294, SEQ ID NO: 295,SEQ ID NO: 296, SEQ ID NO: 297, SEQ ID NO: 298, SEQ ID NO: 299, SEQ IDNO: 300, SEQ ID NO: 301, SEQ ID NO: 302, SEQ ID NO: 303, SEQ ID NO: 304,SEQ ID NO: 305, SEQ ID NO: 306, SEQ ID NO: 307, SEQ ID NO: 308, SEQ IDNO: 309. SEQ ID NO: 310, SEQ ID NO: 321, SEQ ID NO: 322, SEQ ID NO: 323,SEQ ID NO: 324, SEQ ID NO: 325, SEQ ID NO: 326, SEQ ID NO: 327, SEQ IDNO: 328, SEQ ID NO: 329, SEQ ID NO: 330, SEQ ID NO: 335, SEQ ID NO: 338,SEQ ID NO: 339, SEQ ID NO: 341, SEQ ID NO: 342, SEQ ID NO: 343, SEQ IDNO: 344, SEQ ID NO: 345, SEQ ID NO: 346, SEQ ID NO: 347, SEQ ID NO: 348,SEQ ID NO: 349, SEQ ID NO: 350, SEQ ID NO: 351, SEQ ID NO: 352, SEQ IDNO: 353, SEQ ID NO: 354, SEQ ID NO: 355, SEQ ID NO: 357, SEQ ID NO: 358,SEQ ID NO: 359, SEQ ID NO: 360, SEQ ID NO: 362, SEQ ID NO: 364, SEQ IDNO: 366, SEQ ID NO: 367, SEQ ID NO: 369, SEQ ID NO: 370, SEQ ID NO: 371,SEQ ID NO: 372, SEQ ID NO: 377, SEQ ID NO: 379, SEQ ID NO: 380, SEQ IDNO: 382, SEQ ID NO: 385, SEQ ID NO: 388, SEQ ID NO: 389, SEQ ID NO: 391,SEQ ID NO: 392, SEQ ID NO: 393, SEQ ID NO: 394, SEQ ID NO: 396, SEQ IDNO: 397, SEQ ID NO: 398, SEQ ID NO: 399, SEQ ID NO: 400, SEQ ID NO: 401,SEQ ID NO: 402, SEQ ID NO: 403, SEQ ID NO: 404, SEQ ID NO: 405, SEQ IDNO: 406, SEQ ID NO: 407, SEQ ID NO: 408, SEQ ID NO: 409, SEQ ID NO: 410,SEQ ID NO: 411, SEQ ID NO: 413 or SEQ ID NO: 414. “Sufficientlyhomologous” is used herein to refer to an amino acid or nucleic acidsequence that has at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or greater sequence homology compared to a referencesequence using one of the alignment programs described herein usingstandard parameters. One of skill in the art will recognize that thesevalues can be appropriately adjusted to determine corresponding homologyof proteins encoded by two nucleic acid sequences by consideringdegeneracy, amino acid similarity, reading frame positioning, and thelike. In some embodiments, the sequence homology is against thefull-length sequence of the polynucleotide encoding an IPD113polypeptide or against the full-length sequence of an IPD113polypeptide.

In some embodiments, the nucleic acid encodes an IPD113 polypeptidehaving at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or greater sequence identity compared to SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO:255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO:264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO:273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO:311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO:320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO:427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO:436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO:450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO:469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO:480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO:489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 orSEQ ID NO: 495. In some embodiments, the sequence identity is calculatedusing ClustalW algorithm in the ALIGNX® module of the Vector NTI®Program Suite (Invitrogen Corporation, Carlsbad, Calif.) with alldefault parameters. In some embodiments, the sequence identity is acrossthe entire length of polypeptide calculated using ClustalW algorithm inthe ALIGNX module of the Vector NTI Program Suite (InvitrogenCorporation, Carlsbad, Calif.) with all default parameters.

To determine the percent identity of two or more amino acid sequences orof two or more nucleic acid sequences, the sequences are aligned foroptimal comparison purposes. The percent identity between the twosequences is a function of the number of identical positions shared bythe sequences (i.e., percent identity=number of identicalpositions/total number of positions (e.g., overlapping positions)×100).In one embodiment, the two sequences are the same length. In anotherembodiment, the comparison is across the entirety of the referencesequence (e.g., across the entirety of SEQ ID NO: 16). The percentidentity between two sequences can be determined using techniquessimilar to those described below, with or without allowing gaps. Incalculating percent identity, typically exact matches are counted.

Another non-limiting example of a mathematical algorithm utilized forthe comparison of sequences is the algorithm of Needleman and Wunsch,(1970) J. Mol. Biol. 48(3):443-453, used GAP Version 10 software todetermine sequence identity or similarity using the following defaultparameters: % identity and % similarity for a nucleic acid sequenceusing GAP Weight of 50 and Length Weight of 3, and the nwsgapdna.cmpiiscoring matrix; % identity or % similarity for an amino acid sequenceusing GAP weight of 8 and length weight of 2, and the BLOSUM62 scoringprogram. Equivalent programs may also be used. “Equivalent program” isused herein to refer to any sequence comparison program that, for anytwo sequences in question, generates an alignment having identicalnucleotide residue matches and an identical percent sequence identitywhen compared to the corresponding alignment generated by GAP Version10.

In some embodiments, the IPD113 polynucleotide encodes an IPD113polypeptide comprising an amino acid sequence having at least about 80%,81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or greater identity across the entire length ofthe amino acid sequence of SEQ ID NO: 16.

In some embodiments, polynucleotides are provided encoding chimericpolypeptides comprising regions of at least two different IPD113polypeptides of the disclosure.

In some embodiments, polynucleotides are provided encoding chimericpolypeptides comprising regions of at least two different IPD113polypeptides selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495.

In some embodiments, polynucleotides are provided encoding chimericpolypeptides comprising an N-terminal Region of a first IPD113polypeptide of the disclosure operably fused to a C-terminal Region of asecond IPD113 polypeptide of the disclosure.

In some embodiments, polynucleotides are provided encoding chimericpolypeptides comprising an N-terminal Region of a first IPD113polypeptide operably fused to a C-terminal Region of a second IPD113polypeptide, where the IPD113 polypeptide is selected from SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6,SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11,SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ IDNO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30,SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO:35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ IDNO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO:54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ IDNO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68,SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO:73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ IDNO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO:92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ IDNO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ IDNO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110,SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ IDNO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119,SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ IDNO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254,SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ IDNO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263,SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ IDNO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272,SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ IDNO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281,SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ IDNO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319,SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ IDNO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426,SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ IDNO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435,SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ IDNO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448,SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ IDNO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466,SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ IDNO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479,SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ IDNO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488,SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ IDNO: 494 or SEQ ID NO: 495.

In some embodiments, an IPD113 polynucleotide encodes the IPD113polypeptide comprising an amino acid sequence of SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO:255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO:264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO:273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO:311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO:320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO:427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO:436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO:450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO:469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO:480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO:489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 orSEQ ID NO: 495.

The embodiments also encompass nucleic acid molecules encoding IPD113polypeptide variants. “Variants” of the IPD113 polypeptide encodingnucleic acid sequences include those sequences that encode the IPD113polypeptides disclosed herein but that differ conservatively because ofthe degeneracy of the genetic code as well as those that aresufficiently identical as discussed above. Naturally occurring allelicvariants can be identified with the use of molecular biology techniques,such as polymerase chain reaction (PCR) and hybridization techniques asoutlined below. Variant nucleic acid sequences also includesynthetically derived nucleic acid sequences that have been generated,for example, by using site-directed mutagenesis but which still encodethe IPD113 polypeptides disclosed as discussed below.

The present disclosure provides isolated or recombinant polynucleotidesthat encode any of the IPD113 polypeptides disclosed herein. Thosehaving ordinary skill in the art will readily appreciate that due to thedegeneracy of the genetic code, a multitude of nucleotide sequencesencoding IPD113 polypeptides of the present disclosure exist.

The skilled artisan will further appreciate that changes can beintroduced by mutation of the nucleic acid sequences thereby leading tochanges in the amino acid sequence of the encoded IPD113 polypeptides,without altering the biological activity of the proteins. Thus, variantnucleic acid molecules can be created by introducing one or morenucleotide substitutions, additions and/or deletions into thecorresponding nucleic acid sequence disclosed herein, such that one ormore amino acid substitutions, additions or deletions are introducedinto the encoded protein. Mutations can be introduced by standardtechniques, such as site-directed mutagenesis and PCR-mediatedmutagenesis. Such variant nucleic acid sequences are also encompassed bythe present disclosure.

Alternatively, variant nucleic acid sequences can be made by introducingmutations randomly along all or part of the coding sequence, such as bysaturation mutagenesis, and the resultant mutants can be screened forability to confer pesticidal activity to identify mutants that retainactivity. Following mutagenesis, the encoded protein can be expressedrecombinantly, and the activity of the protein can be determined usingstandard assay techniques.

The polynucleotides of the disclosure and fragments thereof areoptionally used as substrates for a variety of recombination andrecursive recombination reactions, in addition to standard cloningmethods as set forth in, e.g., Ausubel, Berger and Sambrook, i.e., toproduce additional pesticidal polypeptide homologues and fragmentsthereof with desired properties. Methods for producing a variant of anynucleic acid listed herein comprising recursively recombining suchpolynucleotide with a second (or more) polynucleotide, thus forming alibrary of variant polynucleotides are also embodiments of thedisclosure, as are the libraries produced, the cells comprising thelibraries and any recombinant polynucleotide produced by such methods.Additionally, such methods optionally comprise selecting a variantpolynucleotide from such libraries based on pesticidal activity, as iswherein such recursive recombination is done in vitro or in vivo.

A variety of diversity generating protocols, including nucleic acidrecursive recombination protocols can be used separately, and/or incombination to produce one or more variants of a nucleic acid or set ofnucleic acids, as well as variants of encoded proteins. Individually andcollectively, these procedures provide robust, widely applicable ways ofgenerating diversified nucleic acids and sets of nucleic acids(including, e.g., nucleic acid libraries) useful, e.g., for theengineering or rapid evolution of nucleic acids, proteins, pathways,cells and/or organisms with new and/or improved characteristics.

While distinctions and classifications are made during the ensuingdiscussion for clarity, it will be appreciated that the techniques areoften not mutually exclusive. Indeed, the various methods can be usedsingly or in combination, in parallel or in series, to access diversesequence variants.

The result of any of the diversity generating procedures describedherein can be the generation of one or more nucleic acids, which can beselected or screened for nucleic acids with or which confer desirableproperties or that encode proteins with or which confer desirableproperties. Following diversification by one or more of the methodsherein or otherwise available to one of skill, any nucleic acids thatare produced can be selected for a desired activity or property, e.g.pesticidal activity or, such activity at a desired pH, etc. This caninclude identifying any activity that can be detected, for example, inan automated or automatable format, by any of the assays in the art,see, e.g., discussion of screening of insecticidal activity, infra. Avariety of related (or even unrelated) properties can be evaluated, inserial or in parallel, at the discretion of the practitioner.

Descriptions of a variety of diversity generating procedures forgenerating modified nucleic acid sequences, e.g., those coding forpolypeptides having pesticidal activity or fragments thereof, are foundin the following publications and the references cited therein: Soong,et al., (2000) Nat Genet 25(4):436-439; Stemmer, et al., (1999) TumorTargeting 4:1-4; Ness, et al., (1999) Nat Biotechnol 17:893-896; Chang,et al., (1999) Nat Biotechnol 17:793-797; Minshull and Stemmer, (1999)Curr Opin Chem Biol 3:284-290; Christians, et al., (1999) Nat Biotechnol17:259-264; Crameri, et al., (1998) Nature 391:288-291; Crameri, et al.,(1997) Nat Biotechnol 15:436-438; Zhang, et al., (1997) PNAS USA94:4504-4509; Patten, et al., (1997) Curr Opin Biotechnol 8:724-733;Crameri, et al., (1996) Nat Med 2:100-103; Crameri, et al., (1996) NatBiotechnol 14:315-319; Gates, et al., (1996) J Mol Biol 255:373-386;Stemmer, (1996) “Sexual PCR and Assembly PCR” In: The Encyclopedia ofMolecular Biology. VCH Publishers, New York. pp. 447-457; Crameri andStemmer, (1995) BioTechniques 18:194-195; Stemmer, et al., (1995) Gene,164:49-53; Stemmer, (1995) Science 270: 1510; Stemmer, (1995)Bio/Technology 13:549-553; Stemmer, (1994) Nature 370:389-391 andStemmer, (1994) PNAS USA 91:10747-10751.

Mutational methods of generating diversity include, for example,site-directed mutagenesis (Ling, et al., (1997) Anal Biochem254(2):157-178; Dale, et al., (1996) Methods Mol Biol 57:369-374; Smith,(1985) Ann Rev Genet 19:423-462; Botstein and Shortle, (1985) Science229:1193-1201; Carter, (1986) Biochem J 237:1-7 and Kunkel, (1987) “Theefficiency of oligonucleotide directed mutagenesis” in Nucleic Acids &Molecular Biology (Eckstein and Lilley, eds., Springer Verlag, Berlin));mutagenesis using uracil containing templates (Kunkel, (1985) PNAS USA82:488-492; Kunkel, et al., (1987) Methods Enzymol 154:367-382 and Bass,et al., (1988) Science 242:240-245); oligonucleotide-directedmutagenesis (Zoller and Smith, (1983) Methods Enzymol 100:468-500;Zoller and Smith, (1987) Methods Enzymol 154:329-350 (1987); Zoller andSmith, (1982) Nucleic Acids Res 10:6487-6500), phosphorothioate-modifiedDNA mutagenesis (Taylor, et al., (1985) Nucl Acids Res 13:8749-8764;Taylor, et al., (1985) Nucl Acids Res 13:8765-8787 (1985); Nakamaye andEckstein, (1986) Nucl Acids Res 14:9679-9698; Sayers, et al., (1988)Nucl Acids Res 16:791-802 and Sayers, et al., (1988) Nucl Acids Res16:803-814); mutagenesis using gapped duplex DNA (Kramer, et al., (1984)Nucl Acids Res 12:9441-9456; Kramer and Fritz, (1987) Methods Enzymol154:350-367; Kramer, et al., (1988) Nucl Acids Res 16:7207 and Fritz, etal., (1988) Nucl Acids Res 16:6987-6999).

Additional suitable methods include point mismatch repair (Kramer, etal., (1984) Cell 38:879-887), mutagenesis using repair-deficient hoststrains (Carter, et al., (1985) Nucl Acids Res 13:4431-4443 and Carter,(1987) Methods in Enzymol 154:382-403), deletion mutagenesis(Eghtedarzadeh and Henikoff, (1986) Nucl Acids Res 14:5115),restriction-selection and restriction-purification (Wells, et al.,(1986) Phil Trans R Soc Lond A 317:415-423), mutagenesis by total genesynthesis (Nambiar, et al., (1984) Science 223:1299-1301; Sakamar andKhorana, (1988) Nucl Acids Res 14:6361-6372; Wells, et al., (1985) Gene34:315-323 and Grundstrom, et al., (1985) Nucl Acids Res 13:3305-3316),double-strand break repair (Mandecki, (1986) PNAS USA, 83:7177-7181 andArnold, (1993) Curr Opin Biotech 4:450-455). Additional details on manyof the above methods can be found in Methods Enzymol Volume 154, whichalso describes useful controls for trouble-shooting problems withvarious mutagenesis methods.

Additional details regarding various diversity generating methods can befound in the following US Patents, PCT Publications and Applications andEPO publications: U.S. Pat. No. 5,723,323, U.S. Pat. No. 5,763,192, U.S.Pat. No. 5,814,476, U.S. Pat. No. 5,817,483, U.S. Pat. No. 5,824,514,U.S. Pat. No. 5,976,862, U.S. Pat. No. 5,605,793, U.S. Pat. No.5,811,238, U.S. Pat. No. 5,830,721, U.S. Pat. No. 5,834,252, U.S. Pat.No. 5,837,458, WO 1995/22625, WO 1996/33207, WO 1997/20078, WO1997/35966, WO 1999/41402, WO 1999/41383, WO 1999/41369, WO 1999/41368,EP 752008, EP 0932670, WO 1999/23107, WO 1999/21979, WO 1998/31837, WO1998/27230, WO 1998/27230, WO 2000/00632, WO 2000/09679, WO 1998/42832,WO 1999/29902, WO 1998/41653, WO 1998/41622, WO 1998/42727, WO2000/18906, WO 2000/04190, WO 2000/42561, WO 2000/42559, WO 2000/42560,WO 2001/23401 and PCT/US01/06775.

The nucleotide sequences of the embodiments can also be used to isolatecorresponding sequences from a fern, including but not limited to aLycopodium species, Huperzia species, and Phlegmariurus species. In thismanner, methods such as PCR, hybridization, and the like can be used toidentify such sequences based on their sequence homology to thesequences set forth herein. Sequences that are selected based on theirsequence identity to the entire sequences set forth herein or tofragments thereof are encompassed by the embodiments. Such sequencesinclude sequences that are orthologs of the disclosed sequences. Theterm “orthologs” refers to genes derived from a common ancestral geneand which are found in different species as a result of speciation.Genes found in different species are considered orthologs when theirnucleotide sequences and/or their encoded protein sequences sharesubstantial identity as defined elsewhere herein. Functions of orthologsare often highly conserved among species.

In a PCR approach, oligonucleotide primers can be designed for use inPCR reactions to amplify corresponding DNA sequences from cDNA orgenomic DNA extracted from any organism of interest. Methods fordesigning PCR primers and PCR cloning are generally known in the art andare disclosed in Sambrook, et al., (1989) Molecular Cloning: ALaboratory Manual (2d ed., Cold Spring Harbor Laboratory Press,Plainview, New York), hereinafter “Sambrook”. See also, Innis, et al.,eds. (1990) PCR Protocols: A Guide to Methods and Applications (AcademicPress, New York); Innis and Gelfand, eds. (1995) PCR Strategies(Academic Press, New York); and Innis and Gelfand, eds. (1999) PCRMethods Manual (Academic Press, New York). Known methods of PCR include,but are not limited to, methods using paired primers, nested primers,single specific primers, degenerate primers, gene-specific primers,vector-specific primers, partially-mismatched primers, and the like.

To identify potential IPD113 polypeptides from fern or other primitiveplants, the fern or other primitive plant cell lysates can be screenedwith antibodies generated against an IPD113 polypeptides and/or IPD113polypeptides using Western blotting and/or ELISA methods. This type ofassays can be performed in a high throughput fashion. Positive samplescan be further analyzed by various techniques such as antibody basedprotein purification and identification.

Alternatively, mass spectrometry based protein identification method canbe used to identify homologs of IPD113 polypeptides using protocols suchas LC-MS/MS based protein identification method is used to associate theMS data of given cell lysate or desired molecular weight enrichedsamples (excised from SDS-PAGE gel of relevant molecular weight bands toIPD113 polypeptides) with sequence information of SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO:255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO:264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO:273, SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO:311, SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQID NO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO:320, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO:427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO:436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO:450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO:469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO:480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO:489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 orSEQ ID NO: 495, and their homologs. Any match in peptide sequencesindicates the potential of having the homologous proteins in thesamples. Additional techniques (protein purification and molecularbiology) can be used to isolate the protein and identify the sequencesof the homologs.

In hybridization methods, all or part of the pesticidal nucleic acidsequence can be used to screen cDNA or genomic libraries. The so-calledhybridization probes may be genomic DNA fragments, cDNA fragments, RNAfragments or other oligonucleotides and may be labeled with a detectablegroup such as 32P or any other detectable marker, such as otherradioisotopes, a fluorescent compound, an enzyme or an enzyme co-factor.Probes for hybridization can be made by labeling syntheticoligonucleotides based on the known IPD113 polypeptide-encoding nucleicacid sequence disclosed herein. Degenerate primers designed based onconserved nucleotides or amino acid residues in the nucleic acidsequence or encoded amino acid sequence can additionally be used. Theprobe typically comprises a region of nucleic acid sequence thathybridizes under stringent conditions to at least about 12, at leastabout 25, at least about 50, 75, 100, 125, 150, 175 or 200 consecutivenucleotides of nucleic acid sequence encoding an IPD113 polypeptide ofthe disclosure or a fragment or variant thereof.

For example, an entire nucleic acid sequence, encoding an IPD113polypeptide, disclosed herein or one or more portions thereof may beused as a probe capable of specifically hybridizing to correspondingnucleic acid sequences encoding IPD113 polypeptide-like sequences andmessenger RNAs. To achieve specific hybridization under a variety ofconditions, such probes include sequences that are unique and arepreferably at least about 10 nucleotides in length or at least about 20nucleotides in length. Such probes may be used to amplify correspondingpesticidal sequences from a chosen organism by PCR. This technique maybe used to isolate additional coding sequences from a desired organismor as a diagnostic assay to determine the presence of coding sequencesin an organism. Hybridization techniques include hybridization screeningof plated DNA libraries (either plaques or colonies; see, for example,Sambrook, et al., (1989) Molecular Cloning: A Laboratory Manual (2d ed.,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).

Hybridization of such sequences may be carried out under stringentconditions. “Stringent conditions” or “stringent hybridizationconditions” is used herein to refer to conditions under which a probewill hybridize to its target sequence to a detectably greater degreethan to other sequences (e.g., at least 2-fold over background).Stringent conditions are sequence-dependent and will be different indifferent circumstances. By controlling the stringency of thehybridization and/or washing conditions, target sequences that are 100%complementary to the probe can be identified (homologous probing).Alternatively, stringency conditions can be adjusted to allow somemismatching in sequences so that lower degrees of similarity aredetected (heterologous probing). Generally, a probe is less than about1000 nucleotides in length, preferably less than 500 nucleotides inlength

Compositions

Compositions comprising at least one IPD113 polypeptide or IPD113chimeric polypeptide of the disclosure are also embraced.

Antibodies

Antibodies to an IPD113 polypeptide of the embodiments or to variants orfragments thereof are also encompassed. The antibodies of the disclosureinclude polyclonal and monoclonal antibodies as well as fragmentsthereof which retain their ability to bind to an IPD113 polypeptidefound in the insect gut. An antibody, monoclonal antibody or fragmentthereof is said to be capable of binding a molecule if it is capable ofspecifically reacting with the molecule to thereby bind the molecule tothe antibody, monoclonal antibody or fragment thereof. The term“antibody” (Ab) or “monoclonal antibody” (Mab) is meant to includeintact molecules as well as fragments or binding regions or domainsthereof (such as, for example, Fab and F(ab).sub.2 fragments) which arecapable of binding hapten. Such fragments are typically produced byproteolytic cleavage, such as papain or pepsin. Alternatively,hapten-binding fragments can be produced through the application ofrecombinant DNA technology or through synthetic chemistry.

A kit for detecting the presence of an IPD113 polypeptide or detectingthe presence of a nucleotide sequence encoding an IPD113 polypeptide ina sample is provided. In one embodiment, the kit provides antibody-basedreagents for detecting the presence of an IPD113 polypeptide in a tissuesample. In another embodiment, the kit provides labeled nucleic acidprobes useful for detecting the presence of one or more polynucleotidesencoding an IPD113 polypeptide. The kit is provided along withappropriate reagents and controls for carrying out a detection method,as well as instructions for use of the kit.

Receptor Identification and Isolation

Receptors to the IPD113 polypeptide of the embodiments or to variants orfragments thereof are also encompassed. Methods for identifyingreceptors can be employed to identify and isolate the receptor thatrecognizes the IPD113 polypeptide using the brush-border membranevesicles from susceptible insects. In addition to the radioactivelabeling method listed in the cited literatures, an IPD113 polypeptidecan be labeled with fluorescent dye and other common labels such asstreptavidin. Brush-border membrane vesicles (BBMV) of susceptibleinsects such as soybean looper and stink bugs can be prepared accordingto the protocols listed in the references and separated on SDS-PAGE geland blotted on suitable membrane. Labeled IPD113 polypeptide can beincubated with blotted membrane of BBMV and labeled IPD113 polypeptidecan be identified with the labeled reporters. Identification of proteinband(s) that interact with the IPD113 polypeptide can be detected byN-terminal amino acid gas phase sequencing or mass spectrometry basedprotein identification method (Patterson, (1998) 10.22, 1-24, CurrentProtocol in Molecular Biology published by John Wiley & Son Inc). Oncethe protein is identified, the corresponding gene can be cloned fromgenomic DNA or cDNA library of the susceptible insects and bindingaffinity can be measured directly with the IPD113 polypeptide. Receptorfunction for insecticidal activity by the IPD113 polypeptide can beverified by accomplished by RNAi type of gene knock out method(Rajagopal, et al., (2002) J. Biol. Chem. 277:46849-46851).

Nucleotide Constructs, Expression Cassettes and Vectors

The use of the term “nucleotide constructs” herein is not intended tolimit the embodiments to nucleotide constructs comprising DNA. Those ofordinary skill in the art will recognize that nucleotide constructsparticularly polynucleotides and oligonucleotides composed ofribonucleotides and combinations of ribonucleotides anddeoxyribonucleotides may also be employed in the methods disclosedherein. The nucleotide constructs, nucleic acids, and nucleotidesequences of the embodiments additionally encompass all complementaryforms of such constructs, molecules, and sequences. Further, thenucleotide constructs, nucleotide molecules, and nucleotide sequences ofthe embodiments encompass all nucleotide constructs, molecules, andsequences which can be employed in the methods of the embodiments fortransforming plants including, but not limited to, those comprised ofdeoxyribonucleotides, ribonucleotides, and combinations thereof. Suchdeoxyribonucleotides and ribonucleotides include both naturallyoccurring molecules and synthetic analogues. The nucleotide constructs,nucleic acids, and nucleotide sequences of the embodiments alsoencompass all forms of nucleotide constructs including, but not limitedto, single-stranded forms, double-stranded forms, hairpins,stem-and-loop structures and the like.

A further embodiment relates to a transformed organism such as anorganism selected from plant and insect cells, bacteria, yeast,baculovirus, protozoa, nematodes and algae. The transformed organismcomprises a DNA molecule of the embodiments, an expression cassettecomprising the DNA molecule or a vector comprising the expressioncassette, which may be stably incorporated into the genome of thetransformed organism.

The sequences of the embodiments are provided in DNA constructs forexpression in the organism of interest. The construct will include 5′and 3′ regulatory sequences operably linked to a sequence of theembodiments. The term “operably linked” as used herein refers to afunctional linkage between a promoter and a second sequence, wherein thepromoter sequence initiates and mediates transcription of the DNAsequence corresponding to the second sequence. Generally, operablylinked means that the nucleic acid sequences being linked are contiguousand where necessary to join two protein coding regions in the samereading frame. The construct may additionally contain at least oneadditional gene to be cotransformed into the organism. Alternatively,the additional gene(s) can be provided on multiple DNA constructs.

Such a DNA construct is provided with a plurality of restriction sitesfor insertion of the IPD113 polypeptide gene sequence of the disclosureto be under the transcriptional regulation of the regulatory regions.The DNA construct may additionally contain selectable marker genes.

The DNA construct will generally include in the 5′ to 3′ direction oftranscription: a transcriptional and translational initiation region(i.e., a promoter), a DNA sequence of the embodiments, and atranscriptional and translational termination region (i.e., terminationregion) functional in the organism serving as a host. Thetranscriptional initiation region (i.e., the promoter) may be native,analogous, foreign or heterologous to the host organism and/or to thesequence of the embodiments. Additionally, the promoter may be thenatural sequence or alternatively a synthetic sequence. The term“foreign” as used herein indicates that the promoter is not found in thenative organism into which the promoter is introduced. Where thepromoter is “foreign” or “heterologous” to the sequence of theembodiments, it is intended that the promoter is not the native ornaturally occurring promoter for the operably linked sequence of theembodiments. As used herein, a chimeric gene comprises a coding sequenceoperably linked to a transcription initiation region that isheterologous to the coding sequence. Where the promoter is a native ornatural sequence, the expression of the operably linked sequence isaltered from the wild-type expression, which results in an alteration inphenotype.

In some embodiments, the DNA construct comprises a polynucleotideencoding an IPD113 polypeptide of the embodiments.

In some embodiments, the DNA construct comprises a polynucleotideencoding a chimeric IPD113 polypeptide of the embodiments.

In some embodiments, the DNA construct comprises a polynucleotideencoding a fusion protein comprising an IPD113 polypeptide of theembodiments.

In some embodiments, the DNA construct comprises a polynucleotidecomprising a first coding sequence encoding the N-terminal Region of afirst IPD113 polypeptide of the disclosure and a second coding sequenceencoding the C-terminal Region of a second IPD113 polypeptide of thedisclosure.

In some embodiments, the DNA construct may also include atranscriptional enhancer sequence. As used herein, the term an“enhancer” refers to a DNA sequence which can stimulate promoteractivity, and may be an innate element of the promoter or a heterologouselement inserted to enhance the level or tissue-specificity of apromoter. Various enhancers including for example, introns with geneexpression enhancing properties in plants (US Patent ApplicationPublication Number 2009/0144863, the ubiquitin intron (i.e., the maizeubiquitin intron 1 (see, for example, NCBI sequence S94464)), the omegaenhancer or the omega prime enhancer (Gallie, et al., (1989) MolecularBiology of RNA ed. Cech (Liss, New York) 237-256 and Gallie, et al.,(1987) Gene 60:217-25), the CaMV 35S enhancer (see, e.g., Benfey, etal., (1990) EMBO J. 9:1685-96) and the enhancers of U.S. Pat. No.7,803,992 may also be used, each of which is incorporated by reference.The above list of transcriptional enhancers is not meant to be limiting.Any appropriate transcriptional enhancer can be used in the embodiments.

The termination region may be native with the transcriptional initiationregion, may be native with the operably linked DNA sequence of interest,may be native with the plant host or may be derived from another source(i.e., foreign or heterologous to the promoter, the sequence ofinterest, the plant host or any combination thereof).

Convenient termination regions are available from the Ti-plasmid of A.tumefaciens, such as the octopine synthase and nopaline synthasetermination regions. See also, Guerineau, et al., (1991) Mol. Gen.Genet. 262:141-144; Proudfoot, (1991) Cell 64:671-674; Sanfacon, et al.,(1991) Genes Dev. 5:141-149; Mogen, et al., (1990) Plant Cell2:1261-1272; Munroe, et al., (1990) Gene 91:151-158; Ballas, et al.,(1989) Nucleic Acids Res. 17:7891-7903 and Joshi, et al., (1987) NucleicAcid Res. 15:9627-9639.

Where appropriate, a nucleic acid may be optimized for increasedexpression in the host organism. Thus, where the host organism is aplant, the synthetic nucleic acids can be synthesized usingplant-preferred codons for improved expression. See, for example,Campbell and Gowri, (1990) Plant Physiol. 92:1-11 for a discussion ofhost-preferred usage. For example, although nucleic acid sequences ofthe embodiments may be expressed in both monocotyledonous anddicotyledonous plant species, sequences can be modified to account forthe specific preferences and GC content preferences of monocotyledons ordicotyledons as these preferences have been shown to differ (Murray etal. (1989) Nucleic Acids Res. 17:477-498). Thus, the maize-preferredcodon for a particular amino acid may be derived from known genesequences from maize. Maize usage for 28 genes from maize plants islisted in Table 4 of Murray, et al., supra. Methods are available in theart for synthesizing plant-preferred genes. See, for example, Murray, etal., (1989) Nucleic Acids Res. 17:477-498, and Liu H et al. Mol Bio Rep37:677-684, 2010, herein incorporated by reference. A Zea maize usagetable can be also found at kazusa.or.jp//cgi-bin/show.cgi?species=4577,which can be accessed using the www prefix.

A Glycine max usage table can be found atkazusa.or.jp//cgi-bin/show.cgi?species=3847&aa=1&style=N, which can beaccessed using the www prefix.

In some embodiments, the recombinant nucleic acid molecule encoding anIPD113 polypeptide has maize optimized codons.

Additional sequence modifications can enhance gene expression in acellular host. These include elimination of sequences encoding spuriouspolyadenylation signals, exon-intron splice site signals,transposon-like repeats, and other well-characterized sequences that maybe deleterious to gene expression. The GC content of the sequence may beadjusted to levels average for a given cellular host, as calculated byreference to known genes expressed in the host cell. The term “hostcell” as used herein refers to a cell which contains a vector andsupports the replication and/or expression of the expression vector isintended. Host cells may be prokaryotic cells such as E. coli oreukaryotic cells such as yeast, insect, amphibian or mammalian cells ormonocotyledonous or dicotyledonous plant cells. An example of amonocotyledonous host cell is a maize host cell. When possible, thesequence is modified to avoid predicted hairpin secondary mRNAstructures.

The expression cassettes may additionally contain 5′ leader sequences.Such leader sequences can act to enhance translation. Translationleaders include: picornavirus leaders, for example, EMCV leader(Encephalomyocarditis 5′ noncoding region) (Elroy-Stein, et al., (1989)Proc. Natl. Acad. Sci. USA 86:6126-6130); potyvirus leaders, forexample, TEV leader (Tobacco Etch Virus) (Gallie, et al., (1995) Gene165(2):233-238), MDMV leader (Maize Dwarf

Mosaic Virus), human immunoglobulin heavy-chain binding protein (BiP)(Macejak, et al., (1991) Nature 353:90-94); untranslated leader from thecoat protein mRNA of alfalfa mosaic virus (AMV RNA 4) (Jobling, et al.,(1987) Nature 325:622-625); tobacco mosaic virus leader (TMV) (Gallie,et al., (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York),pp. 237-256) and maize chlorotic mottle virus leader (MCMV) (Lommel, etal., (1991) Virology 81:382-385). See also, Della-Cioppa, et al., (1987)Plant Physiol. 84:965-968. Such constructs may also contain a “signalsequence” or “leader sequence” to facilitate co-translational orpost-translational transport of the peptide to certain intracellularstructures such as the chloroplast (or other plastid), endoplasmicreticulum or Golgi apparatus.

“Signal sequence” as used herein refers to a sequence that is known orsuspected to result in cotranslational or post-translational peptidetransport across the cell membrane. In eukaryotes, this typicallyinvolves secretion into the Golgi apparatus, with some resultingglycosylation. Insecticidal toxins of bacteria are often synthesized asprotoxins, which are proteolytically activated in the gut of the targetpest (Chang, (1987) Methods Enzymol. 153:507-516). In some embodiments,the signal sequence is in the native sequence or may be derived from asequence of the embodiments. “Leader sequence” as used herein refers toany sequence that when translated, results in an amino acid sequencesufficient to trigger co-translational transport of the peptide chain toa subcellular organelle. Thus, this includes leader sequences targetingtransport and/or glycosylation by passage into the endoplasmicreticulum, passage to vacuoles, plastids including chloroplasts,mitochondria, and the like. Nuclear-encoded proteins targeted to thechloroplast thylakoid lumen compartment have a characteristic bipartitetransit peptide, composed of a stromal targeting signal peptide and alumen targeting signal peptide. The stromal targeting information is inthe amino-proximal portion of the transit peptide. The lumen targetingsignal peptide is in the carboxyl-proximal portion of the transitpeptide, and contains all the information for targeting to the lumen.Recent research in proteomics of the higher plant chloroplast hasachieved in the identification of numerous nuclear-encoded lumenproteins (Kieselbach et al. FEBS LETT480:271-276, 2000; Peltier et al.Plant Cell 12:319-341, 2000; Bricker et al. Biochim. Biophys Acta1503:350-356, 2001), the lumen targeting signal peptide of which canpotentially be used in accordance with the present disclosure. About 80proteins from Arabidopsis, as well as homologous proteins from spinachand garden pea, are reported by Kieselbach et al., PhotosynthesisResearch, 78:249-264, 2003. Table 2 of this publication, which isincorporated into the description herewith by reference, discloses 85proteins from the chloroplast lumen, identified by their accessionnumber (see also US Patent Application Publication 2009/09044298). Inaddition, the recently published draft version of the rice genome (Goffet al, Science 296:92-100, 2002) is a suitable source for lumentargeting signal peptide which may be used in accordance with thepresent disclosure.

Suitable chloroplast transit peptides (CTP) include chimeric CT'scomprising but not limited to, an N-terminal domain, a central domain ora C-terminal domain from a CTP from Oryza sativa 1-decoy-Dxylose-5-Phosphate Synthase Oryza sativa-Superoxide dismutase Oryzasativa-soluble starch synthase Oryza sativa-NADP-dependent Malic acidenzyme Oryza sativa-Phospho-2-dehydro-3-deoxyheptonate Aldolase 2 Oryzasativa-L-Ascorbate peroxidase 5 Oryza sativa-Phosphoglucan waterdikinase, Zea Mays ssRUBISCO, Zea Mays-beta-glucosidase, Zea Mays-Malatedehydrogenase, Zea Mays Thioredoxin M-type US Patent ApplicationPublication 2012/0304336).

The IPD113 polypeptide gene to be targeted to the chloroplast may beoptimized for expression in the chloroplast to account for differencesin usage between the plant nucleus and this organelle. In this manner,the nucleic acids of interest may be synthesized usingchloroplast-preferred sequences.

In preparing the expression cassette, the various DNA fragments may bemanipulated to provide for the DNA sequences in the proper orientationand, as appropriate, in the proper reading frame. Toward this end,adapters or linkers may be employed to join the DNA fragments or othermanipulations may be involved to provide for convenient restrictionsites, removal of superfluous DNA, removal of restriction sites or thelike. For this purpose, in vitro mutagenesis, primer repair,restriction, annealing, resubstitutions, e.g., transitions andtransversions, may be involved.

Several promoters can be used in the practice of the embodiments. Thepromoters can be selected based on the desired outcome. The nucleicacids can be combined with constitutive, tissue-preferred, inducible orother promoters for expression in the host organism. Suitableconstitutive promoters for use in a plant host cell include, forexample, the core promoter of the Rsyn7 promoter and other constitutivepromoters disclosed in WO 1999/43838 and U.S. Pat. No. 6,072,050; thecore CaMV 35S promoter (Odell, et al., (1985) Nature 313:810-812); riceactin (McElroy, et al., (1990) Plant Cell 2:163-171); ubiquitin(Christensen, et al., (1989) Plant Mol. Biol. 12:619-632 andChristensen, et al., (1992) Plant Mol. Biol. 18:675-689); pEMU (Last, etal., (1991) Theor. Appl. Genet. 81:581-588); MAS (Velten, et al., (1984)EMBO J. 3:2723-2730); ALS promoter (U.S. Pat. No. 5,659,026) and thelike. Other constitutive promoters include, for example, those discussedin U.S. Pat. Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785;5,399,680; 5,268,463; 5,608,142 and 6,177,611.

Depending on the desired outcome, it may be beneficial to express thegene from an inducible promoter. Of particular interest for regulatingthe expression of the nucleotide sequences of the embodiments in plantsare wound-inducible promoters. Such wound-inducible promoters, mayrespond to damage caused by insect feeding, and include potatoproteinase inhibitor (pin II) gene (Ryan, (1990) Ann. Rev. Phytopath.28:425-449; Duan, et al., (1996) Nature Biotechnology 14:494-498); wun1and wun2, U.S. Pat. No. 5,428,148; win1 and win2 (Stanford, et al.,(1989) Mol. Gen. Genet. 215:200-208); systemin (McGurl, et al., (1992)Science 225:1570-1573); WIP1 (Rohmeier, et al., (1993) Plant Mol. Biol.22:783-792; Eckelkamp, et al., (1993) FEBS Letters 323:73-76); MPI gene(Corderok, et al., (1994) Plant J. 6(2):141-150) and the like, hereinincorporated by reference.

Additionally, pathogen-inducible promoters may be employed in themethods and nucleotide constructs of the embodiments. Suchpathogen-inducible promoters include those from pathogenesis-relatedproteins (PR proteins), which are induced following infection by apathogen; e.g., PR proteins, SAR proteins, beta-1,3-glucanase,chitinase, etc. See, for example, Redolfi, et al., (1983) Neth. J. PlantPathoL 89:245-254; Uknes, et al., (1992) Plant Cell 4: 645-656 and VanLoon, (1985) Plant Mol. ViroL 4:111-116. See also, WO 1999/43819, hereinincorporated by reference.

Of interest are promoters that are expressed locally at or near the siteof pathogen infection. See, for example, Marineau, et al., (1987) PlantMol. Biol. 9:335-342; Matton, et al., (1989) Molecular Plant-MicrobeInteractions 2:325-331; Somsisch, et al., (1986) Proc. Natl. Acad. Sci.USA 83:2427-2430; Somsisch, et al., (1988) Mol. Gen. Genet. 2:93-98 andYang, (1996) Proc. Natl. Acad. Sci. USA 93:14972-14977. See also, Chen,et al., (1996) Plant J. 10:955-966; Zhang, et al., (1994) Proc. Natl.Acad. Sci. USA 91:2507-2511; Warner, et al., (1993) Plant J. 3:191-201;Siebertz, et al., (1989) Plant Cell 1:961-968; U.S. Pat. No. 5,750,386(nematode-inducible) and the references cited therein. Of particularinterest is the inducible promoter for the maize PRms gene, whoseexpression is induced by the pathogen Fusarium moniliforme (see, forexample, Cordero, et al., (1992) Physiol. Mol. Plant Path. 41:189-200).

Chemical-regulated promoters can be used to modulate the expression of agene in a plant through the application of an exogenous chemicalregulator. Depending upon the objective, the promoter may be achemical-inducible promoter, where application of the chemical inducesgene expression or a chemical-repressible promoter, where application ofthe chemical represses gene expression. Chemical-inducible promotersinclude, but are not limited to, the maize In2-2 promoter, which isactivated by benzenesulfonamide herbicide safeners, the maize GSTpromoter, which is activated by hydrophobic electrophilic compounds thatare used as pre-emergent herbicides, and the tobacco PR-1 a promoter,which is activated by salicylic acid. Other chemical-regulated promotersof interest include steroid-responsive promoters (see, for example, theglucocorticoid-inducible promoter in Schena, et al., (1991) Proc. Natl.Acad. Sci. USA 88:10421-10425 and McNellis, et al., (1998) Plant J.14(2):247-257) and tetracycline-inducible and tetracycline-repressiblepromoters (see, for example, Gatz, et al., (1991) Mol. Gen. Genet.227:229-237 and U.S. Pat. Nos. 5,814,618 and 5,789,156), hereinincorporated by reference.

Tissue-preferred promoters can be utilized to target enhanced an IPD113polypeptide expression within a particular plant tissue.Tissue-preferred promoters include those discussed in Yamamoto, et al.,(1997) Plant J. 12(2)255-265; Kawamata, et al., (1997) Plant CellPhysiol. 38(7):792-803; Hansen, et al., (1997) Mol. Gen Genet.254(3):337-343; Russell, et al., (1997) Transgenic Res. 6(2):157-168;Rinehart, et al., (1996) Plant Physiol. 112(3):1331-1341; Van Camp, etal., (1996) Plant Physiol. 112(2):525-535; Canevascini, et al., (1996)Plant Physiol. 112(2):513-524; Yamamoto, et al., (1994) Plant CellPhysiol. 35(5):773-778; Lam, (1994) Results ProbL Cell Differ.20:181-196; Orozco, et al., (1993) Plant Mol Biol. 23(6):1129-1138;Matsuoka, et al., (1993) Proc Natl. Acad. Sci. USA 90(20):9586-9590 andGuevara-Garcia, et al., (1993) Plant J. 4(3):495-505. Such promoters canbe modified, if necessary, for weak expression.

Leaf-preferred promoters are known in the art. See, for example,Yamamoto, et al., (1997) Plant J. 12(2):255-265; Kwon, et al., (1994)Plant Physiol. 105:357-67; Yamamoto, et al., (1994) Plant Cell Physiol.35(5):773-778; Gotor, et al., (1993) Plant J. 3:509-18; Orozco, et al.,(1993) Plant Mol. Biol. 23(6):1129-1138 and Matsuoka, et al., (1993)Proc. Natl. Acad. Sci. USA 90(20):9586-9590.

Root-preferred or root-specific promoters can be selected from the manyavailable from the literature or isolated de novo from variouscompatible species. See, for example, Hire, et al., (1992) Plant Mol.Biol. 20(2):207-218 (soybean root-specific glutamine synthetase gene);Keller and Baumgartner, (1991) Plant Cell 3(10):1051-1061 (root-specificcontrol element in the GRP 1.8 gene of French bean); Sanger, et al.,(1990) Plant Mol. Biol. 14(3):433-443 (root-specific promoter of themannopine synthase (MAS) gene of Agrobacterium tumefaciens) and Miao, etal., (1991) Plant Cell 3(1):11-22 (full-length cDNA clone encodingcytosolic glutamine synthetase (GS), which is expressed in roots androot nodules of soybean). See also, Bogusz, et al., (1990) Plant Cell2(7):633-641, where two root-specific promoters isolated from hemoglobingenes from the nitrogen-fixing nonlegume Parasponia andersonii and therelated non-nitrogen-fixing nonlegume Trema tomentosa are described. Thepromoters of these genes were linked to a β-glucuronidase reporter geneand introduced into both the nonlegume Nicotiana tabacum and the legumeLotus corniculatus, and in both instances root-specific promoteractivity was preserved. Leach and Aoyagi, (1991) describe their analysisof the promoters of the highly expressed roIC and roID root-inducinggenes of Agrobacterium rhizogenes (see, Plant Science (Limerick)79(1):69-76). They concluded that enhancer and tissue-preferred DNAdeterminants are dissociated in those promoters. Teeri, et al., (1989)used gene fusion to lacZ to show that the Agrobacterium T-DNA geneencoding octopine synthase is especially active in the epidermis of theroot tip and that the TR2′ gene is root specific in the intact plant andstimulated by wounding in leaf tissue, an especially desirablecombination of characteristics for use with an insecticidal orlarvicidal gene (see, EMBO J. 8(2):343-350). The TR1′ gene fused tonptII (neomycin phosphotransferase II) showed similar characteristics.Additional root-preferred promoters include the VfENOD-GRP3 genepromoter (Kuster, et al., (1995) Plant Mol. Biol. 29(4):759-772) androIB promoter (Capana, et al., (1994) Plant Mol. Biol. 25(4):681-691.See also, U.S. Pat. Nos. 5,837,876; 5,750,386; 5,633,363; 5,459,252;5,401,836; 5,110,732 and 5,023,179. Arabidopsis thaliana root-pref erredregulatory sequences are disclosed in US20130117883.

“Seed-preferred” promoters include both “seed-specific” promoters (thosepromoters active during seed development such as promoters of seedstorage proteins) as well as “seed-germinating” promoters (thosepromoters active during seed germination). See, Thompson, et al., (1989)BioEssays 10:108, herein incorporated by reference. Such seed-preferredpromoters include, but are not limited to, Cim1 (cytokinin-inducedmessage); cZ19B1 (maize 19 kDa zein); and milps(myo-inositol-1-phosphate synthase) (see, U.S. Pat. No. 6,225,529,herein incorporated by reference). Gamma-zein and Glb-1 areendosperm-specific promoters. For dicots, seed-specific promotersinclude, but are not limited to, Kunitz trypsin inhibitor 3 (KTi3)(Jofuku and Goldberg, (1989) Plant Cell 1:1079-1093), bean β-phaseolin,napin, β-conglycinin, glycinin 1, soybean lectin, cruciferin, and thelike. For monocots, seed-specific promoters include, but are not limitedto, maize 15 kDa zein, 22 kDa zein, 27 kDa zein, g-zein, waxy, shrunken1, shrunken 2, globulin 1, etc. See also, WO 2000/12733, whereseed-preferred promoters from end1 and end2 genes are disclosed; hereinincorporated by reference. In dicots, seed specific promoters includebut are not limited to seed coat promoter from Arabidopsis, pBAN; andthe early seed promoters from Arabidopsis, p26, p63, and p63tr (U.S.Pat. Nos. 7,294,760 and 7,847,153). A promoter that has “preferred”expression in a particular tissue is expressed in that tissue to agreater degree than in at least one other plant tissue. Sometissue-preferred promoters show expression almost exclusively in theparticular tissue.

Where low level expression is desired, weak promoters will be used.Generally, the term “weak promoter” as used herein refers to a promoterthat drives expression of a coding sequence at a low level. By low levelexpression at levels of between about 1/1000 transcripts to about1/100,000 transcripts to about 1/500,000 transcripts is intended.Alternatively, it is recognized that the term “weak promoters” alsoencompasses promoters that drive expression in only a few cells and notin others to give a total low level of expression. Where a promoterdrives expression at unacceptably high levels, portions of the promotersequence can be deleted or modified to decrease expression levels.

Such weak constitutive promoters include, for example the core promoterof the Rsyn7 promoter (WO 1999/43838 and U.S. Pat. No. 6,072,050), thecore 35S CaMV promoter, and the like. Other constitutive promotersinclude, for example, those disclosed in U.S. Pat. Nos. 5,608,149;5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463;5,608,142 and 6,177,611, herein incorporated by reference.

The above list of promoters is not meant to be limiting. Any appropriatepromoter can be used in the embodiments.

Generally, the expression cassette will comprise a selectable markergene for the selection of transformed cells. Selectable marker genes areutilized for the selection of transformed cells or tissues. Marker genesinclude genes encoding antibiotic resistance, such as those encodingneomycin phosphotransferase II (NEO) and hygromycin phosphotransferase(HPT), as well as genes conferring resistance to herbicidal compounds,such as glufosinate ammonium, bromoxynil, imidazolinones and2,4-dichlorophenoxyacetate (2,4-D). Additional examples of suitableselectable marker genes include, but are not limited to, genes encodingresistance to chloramphenicol (Herrera Estrella, et al., (1983) EMBO J.2:987-992); methotrexate (Herrera Estrella, et al., (1983) Nature303:209-213 and Meijer, et al., (1991) Plant Mol. Biol. 16:807-820);streptomycin (Jones, et al., (1987) Mol. Gen. Genet. 210:86-91);spectinomycin (Bretagne-Sagnard, et al., (1996) Transgenic Res.5:131-137); bleomycin (Hille, et al., (1990) Plant Mol. Biol.7:171-176); sulfonamide (Guerineau, et al., (1990) Plant Mol. Biol.15:127-136); bromoxynil (Stalker, et al., (1988) Science 242:419-423);glyphosate (Shaw, et al., (1986) Science 233:478-481 and U.S. patentapplication Ser. Nos. 10/004,357 and 10/427,692); phosphinothricin(DeBlock, et al., (1987) EMBO J. 6:2513-2518). See generally, Yarranton,(1992) Curr. Opin. Biotech. 3:506-511; Christopherson, et al., (1992)Proc. Natl. Acad. Sci. USA 89:6314-6318; Yao, et al., (1992) Cell71:63-72; Reznikoff, (1992) Mol. Microbiol. 6:2419-2422; Barkley, etal., (1980) in The Operon, pp. 177-220; Hu, et al., (1987) Cell48:555-566; Brown, et al., (1987) Cell 49:603-612; Figge, et al., (1988)Cell 52:713-722; Deuschle, et al., (1989) Proc. Natl. Acad. Sci. USA86:5400-5404; Fuerst, et al., (1989) Proc. Natl. Acad. Sci. USA86:2549-2553; Deuschle, et al., (1990) Science 248:480-483; Gossen,(1993) Ph.D. Thesis, University of Heidelberg; Reines, et al., (1993)Proc. Natl. Acad. Sci. USA 90:1917-1921; Labow, et al., (1990) Mol.Cell. Biol. 10:3343-3356; Zambretti, et al., (1992) Proc. Natl. Acad.Sci. USA 89:3952-3956; Baim, et al., (1991) Proc. Natl. Acad. Sci. USA88:5072-5076; Wyborski, et al., (1991) Nucleic Acids Res. 19:4647-4653;Hillenand-Wissman, (1989) Topics Mol. Struc. Biol. 10:143-162;Degenkolb, et al., (1991) Antimicrob. Agents Chemother. 35:1591-1595;Kleinschnidt, etaL, (1988) Biochemistry 27:1094-1104; Bonin, (1993)Ph.D. Thesis, University of Heidelberg; Gossen, et al., (1992) Proc.Natl. Acad. Sci. USA 89:5547-5551; Oliva, et al., (1992) Antimicrob.Agents Chemother. 36:913-919; Hlavka, et al., (1985) Handbook ofExperimental Pharmacology, Vol. 78 (Springer-Verlag, Berlin) and Gill,et al., (1988) Nature 334:721-724. Such disclosures are hereinincorporated by reference.

The above list of selectable marker genes is not meant to be limiting.Any selectable marker gene can be used in the embodiments.

Plant Transformation

The methods of the embodiments involve introducing a polypeptide orpolynucleotide into a plant. “Introducing” is as used herein meanspresenting to the plant the polynucleotide or polypeptide in such amanner that the sequence gains access to the interior of a cell of theplant. The methods of the embodiments do not depend on a particularmethod for introducing a polynucleotide or polypeptide into a plant,only that the polynucleotide or polypeptides gains access to theinterior of at least one cell of the plant. Methods for introducingpolynucleotide or polypeptides into plants including, but not limitedto, stable transformation methods, transient transformation methods, andvirus-mediated methods.

“Stable transformation” is as used herein means that the nucleotideconstruct introduced into a plant integrates into the genome of theplant and is capable of being inherited by the progeny thereof.“Transient transformation” as used herein means that a polynucleotide isintroduced into the plant and does not integrate into the genome of theplant or a polypeptide is introduced into a plant. “Plant” as usedherein refers to whole plants, plant organs (e.g., leaves, stems, roots,etc.), seeds, plant cells, propagules, embryos and progeny of the same.Plant cells can be differentiated or undifferentiated (e.g. callus,suspension culture cells, protoplasts, leaf cells, root cells, phloemcells and pollen).

Transformation protocols as well as protocols for introducing nucleotidesequences into plants may vary depending on the type of plant or plantcell, i.e., monocot or dicot, targeted for transformation. Suitablemethods of introducing nucleotide sequences into plant cells andsubsequent insertion into the plant genome include microinjection(Crossway, et al., (1986) Biotechniques 4:320-334), electroporation(Riggs, et al., (1986) Proc. Natl. Acad. Sci. USA 83:5602-5606),Agrobacterium-mediated transformation (U.S. Pat. Nos. 5,563,055 and5,981,840), direct gene transfer (Paszkowski, et al., (1984) EMBO J.3:2717-2722) and ballistic particle acceleration (see, for example, U.S.Pat. Nos. 4,945,050; 5,879,918; 5,886,244 and 5,932,782; Tomes, et al.,(1995) in Plant Cell, Tissue, and Organ Culture: Fundamental Methods,ed. Gamborg and Phillips, (Springer-Verlag, Berlin) and McCabe, et al.,(1988) Biotechnology 6:923-926) and Lecl transformation (WO 00/28058).For potato transformation see, Tu, et al., (1998) Plant MolecularBiology 37:829-838 and Chong, et al., (2000) Transgenic Research9:71-78. Additional transformation procedures can be found inWeissinger, et al., (1988) Ann. Rev. Genet. 22:421-477; Sanford, et al.,(1987) Particulate Science and Technology 5:27-37 (onion); Christou, etal., (1988) Plant Physiol. 87:671-674 (soybean); McCabe, et al., (1988)Bio/Technology 6:923-926 (soybean); Finer and McMullen, (1991) In VitroCell Dev. Biol. 27P:175-182 (soybean); Singh, et al., (1998) Theor.Appl. Genet. 96:319-324 (soybean); Datta, et al., (1990) Biotechnology8:736-740 (rice); Klein, et al., (1988) Proc. Natl. Acad. Sci. USA85:4305-4309 (maize); Klein, et al., (1988) Biotechnology 6:559-563(maize); U.S. Pat. Nos. 5,240,855; 5,322,783 and 5,324,646; Klein, etal., (1988) Plant Physiol. 91:440-444 (maize); Fromm, et al., (1990)Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren, et al., (1984)Nature (London) 311:763-764; U.S. Pat. No. 5,736,369 (cereals);Bytebier, et al., (1987) Proc. Natl. Acad. Sci. USA 84:5345-5349(Liliaceae); De Wet, et al., (1985) in The Experimental Manipulation ofOvule Tissues, ed. Chapman, et al., (Longman, New York), pp. 197-209(pollen); Kaeppler, et al., (1990) Plant Cell Reports 9:415-418 andKaeppler, et al., (1992) Theor. Appl. Genet. 84:560-566(whisker-mediated transformation); D'Halluin, et al., (1992) Plant Cell4:1495-1505 (electroporation); Li, et al., (1993) Plant Cell Reports12:250-255 and Christou and Ford, (1995) Annals of Botany 75:407-413(rice); Osjoda, et al., (1996) Nature Biotechnology 14:745-750 (maizevia Agrobacterium tumefaciens); all of which are herein incorporated byreference.

In specific embodiments, the sequences of the embodiments can beprovided to a plant using a variety of transient transformation methods.Such transient transformation methods include, but are not limited to,the introduction of the IPD113 polynucleotide or variants and fragmentsthereof directly into the plant or the introduction of the IPD113polypeptide transcript into the plant. Such methods include, forexample, microinjection or particle bombardment. See, for example,Crossway, et al., (1986) Mol Gen. Genet. 202:179-185; Nomura, et al.,(1986) Plant Sci. 44:53-58; Hepler, et al., (1994) Proc. Natl. Acad.Sci. 91:2176-2180 and Hush, et al., (1994) The Journal of Cell Science107:775-784, all of which are herein incorporated by reference.Alternatively, the IPD113 polynucleotide can be transiently transformedinto the plant. Such techniques include viral vector system and theprecipitation of the polynucleotide in a manner that precludessubsequent release of the DNA. Thus, transcription from theparticle-bound DNA can occur, but the frequency with which it isreleased to become integrated into the genome is greatly reduced. Suchmethods include the use of particles coated with polyethylimine (PEI;Sigma #P3143).

Methods for the targeted insertion of a polynucleotide at a specificlocation in the plant genome can be achieved by the insertion of thepolynucleotide at a desired genomic location is achieved using asite-specific recombination system. See, for example, WO 1999/25821, WO1999/25854, WO 1999/25840, WO 1999/25855 and WO 1999/25853, all of whichare herein incorporated by reference. Briefly, the polynucleotide of theembodiments can be contained in transfer cassette flanked by twonon-identical recombination sites. The transfer cassette is introducedinto a plant have stably incorporated into its genome a target sitewhich is flanked by two non-identical recombination sites thatcorrespond to the sites of the transfer cassette. An appropriaterecombinase is provided and the transfer cassette is integrated at thetarget site. The polynucleotide of interest is thereby integrated at aspecific chromosomal position in the plant genome.

Plant transformation vectors may be comprised of one or more DNA vectorsneeded for achieving plant transformation. For example, it is a commonpractice in the art to utilize plant transformation vectors that arecomprised of more than one contiguous DNA segment.

These vectors are often referred to in the art as “binary vectors”.Binary vectors as well as vectors with helper plasmids are most oftenused for Agrobacterium-mediated transformation, where the size andcomplexity of DNA segments needed to achieve efficient transformation isquite large, and it is advantageous to separate functions onto separateDNA molecules. Binary vectors typically contain a plasmid vector thatcontains the cis-acting sequences required for T-DNA transfer (such asleft border and right border), a selectable marker that is engineered tobe capable of expression in a plant cell, and a “gene of interest” (agene engineered to be capable of expression in a plant cell for whichgeneration of transgenic plants is desired). Also present on thisplasmid vector are sequences required for bacterial replication. Thecis-acting sequences are arranged in a fashion to allow efficienttransfer into plant cells and expression therein. For example, theselectable marker gene and the pesticidal gene are located between theleft and right borders. Often a second plasmid vector contains thetrans-acting factors that mediate T-DNA transfer from Agrobacterium toplant cells. This plasmid often contains the virulence functions (Virgenes) that allow infection of plant cells by Agrobacterium, andtransfer of DNA by cleavage at border sequences and vir-mediated DNAtransfer, as is understood in the art (Hellens and Mullineaux, (2000)Trends in Plant Science 5:446-451). Several types of Agrobacteriumstrains (e.g. LBA4404, GV3101, EHA101, EHA105, etc.) can be used forplant transformation. The second plasmid vector is not necessary fortransforming the plants by other methods such as microprojection,microinjection, electroporation, polyethylene glycol, etc.

In general, plant transformation methods involve transferringheterologous DNA into target plant cells (e.g., immature or matureembryos, suspension cultures, undifferentiated callus, protoplasts,etc.), followed by applying a maximum threshold level of appropriateselection (depending on the selectable marker gene) to recover thetransformed plant cells from a group of untransformed cell mass.Following integration of heterologous foreign DNA into plant cells, onethen applies a maximum threshold level of appropriate selection in themedium to kill the untransformed cells and separate and proliferate theputatively transformed cells that survive from this selection treatmentby transferring regularly to a fresh medium. By continuous passage andchallenge with appropriate selection, one can identify and proliferatethe cells that are transformed with the plasmid vector. Molecular andbiochemical methods can then be used to confirm the presence of theintegrated heterologous gene of interest into the genome of thetransgenic plant.

Explants are typically transferred to a fresh supply of the same mediumand cultured routinely. Subsequently, the transformed cells aredifferentiated into shoots after placing on regeneration mediumsupplemented with a maximum threshold level of selecting agent. Theshoots are then transferred to a selective rooting medium for recoveringrooted shoot or plantlet. The transgenic plantlet then grows into amature plant and produces fertile seeds (e.g., Hiei, et al., (1994) ThePlant Journal 6:271-282; Ishida, et al., (1996) Nature Biotechnology14:745-750). Explants are typically transferred to a fresh supply of thesame medium and cultured routinely. A general description of thetechniques and methods for generating transgenic plants are found inAyres and Park, (1994) Critical Reviews in Plant Science 13:219-239 andBommineni and Jauhar, (1997) Maydica 42:107-120. Since the transformedmaterial contains many cells; both transformed and non-transformed cellsare present in any piece of subjected target callus or tissue or groupof cells. The ability to kill non-transformed cells and allowtransformed cells to proliferate results in transformed plant cultures.Often, the ability to remove non-transformed cells is a limitation torapid recovery of transformed plant cells and successful generation oftransgenic plants.

The cells that have been transformed may be grown into plants inaccordance with conventional ways. See, for example, McCormick, et al.,(1986) Plant Cell Reports 5:81-84. These plants may then be grown, andeither pollinated with the same transformed strain or different strains,and the resulting hybrid having constitutive or inducible expression ofthe desired phenotypic characteristic identified. Two or moregenerations may be grown to ensure that expression of the desiredphenotypic characteristic is stably maintained and inherited and thenseeds harvested to ensure that expression of the desired phenotypiccharacteristic has been achieved.

The nucleotide sequences of the embodiments may be provided to the plantby contacting the plant with a virus or viral nucleic acids. Generally,such methods involve incorporating the nucleotide construct of interestwithin a viral DNA or RNA molecule. It is recognized that therecombinant proteins of the embodiments may be initially synthesized aspart of a viral polyprotein, which later may be processed by proteolysisin vivo or in vitro to produce the desired IPD113 polypeptide. It isalso recognized that such a viral polyprotein, comprising at least aportion of the amino acid sequence of an IPD113 of the embodiments, mayhave the desired pesticidal activity. Such viral polyproteins and thenucleotide sequences that encode for them are encompassed by theembodiments. Methods for providing plants with nucleotide constructs andproducing the encoded proteins in the plants, which involve viral DNA orRNA molecules. See, for example, U.S. Pat. Nos. 5,889,191; 5,889,190;5,866,785; 5,589,367 and 5,316,931; herein incorporated by reference.

Methods for transformation of chloroplasts include, for example, Svab,et al., (1990) Proc. Natl. Acad. Sci. USA 87:8526-8530; Svab and Maliga,(1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab and Maliga, (1993)EMBO J. 12:601-606. The method relies on particle gun delivery of DNAcontaining a selectable marker and targeting of the DNA to the plastidgenome through homologous recombination. Additionally, plastidtransformation can be accomplished by transactivation of a silentplastid-borne transgene by tissue-preferred expression of anuclear-encoded and plastid-directed RNA polymerase. Such a system hasbeen reported in McBride, et al., (1994) Proc. Natl. Acad. Sci. USA91:7301-7305.

The embodiments further relate to plant-propagating material of atransformed plant of the embodiments including, but not limited to,seeds, tubers, corms, bulbs, leaves and cuttings of roots and shoots.

The embodiments may be used for transformation of any plant species,including, but not limited to, monocots and dicots. Examples of plantsof interest include, but are not limited to, corn (Zea mays), Brassicasp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassicaspecies useful as sources of seed oil, alfalfa (Medicago sativa), rice(Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghumvulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet(Panicum miliaceum), foxtail millet (Setaria italica), finger millet(Eleusine coracana)), sunflower (Helianthus annuus), safflower(Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycinemax), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts(Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum),sweet potato (Ipomoea batatus), cassava (Manihot esculenta), coffee(Coffea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus),citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camelliasinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficuscasica), guava (Psidium guajava), mango (Mangifera indica), olive (Oleaeuropaea), papaya (Carica papaya), cashew (Anacardium occidentale),macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugarbeets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley,vegetables ornamentals, and conifers.

Vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g.,Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseoluslimensis), peas (Lathyrus spp.), and members of the genus Cucumis suchas cucumber (C. sativus), cantaloupe (C. cantalupensis), and musk melon(C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea(Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosaspp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias(Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia(Euphorbia pulcherrima), and chrysanthemum. Conifers that may beemployed in practicing the embodiments include, for example, pines suchas loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosapine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Montereypine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Westernhemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood(Sequoia sempervirens); true firs such as silver fir (Abies amabilis)and balsam fir (Abies balsamea); and cedars such as Western red cedar(Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis).Plants of the embodiments include crop plants (for example, corn,alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut,sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants.

Turf grasses include, but are not limited to: annual bluegrass (Poaannua); annual ryegrass (Lolium multiflorum); Canada bluegrass (Poacompressa); Chewing's fescue (Festuca rubra); colonial bentgrass(Agrostis tenuis); creeping bentgrass (Agrostis palustris); crestedwheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyroncristatum); hard fescue (Festuca longifolia); Kentucky bluegrass (Poapratensis); orchardgrass (Dactylis glomerata); perennial ryegrass(Lolium perenne); red fescue (Festuca rubra); redtop (Agrostis alba);rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina); smoothbromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy(Phleum pratense); velvet bentgrass (Agrostis canina); weepingalkaligrass (Puccinellia distans); western wheatgrass (Agropyronsmithii); Bermuda grass (Cynodon spp.); St. Augustine grass(Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass(Paspalum notatum); carpet grass (Axonopus affinis); centipede grass(Eremochloa ophiuroides); kikuyu grass (Pennisetum clandesinum);seashore paspalum (Paspalum vaginatum); blue gramma (Boutelouagracilis); buffalo grass (Buchloe dactyloids); sideoats gramma(Bouteloua curtipendula).

Plants of interest include grain plants that provide seeds of interest,oil-seed plants, and leguminous plants. Seeds of interest include grainseeds, such as corn, wheat, barley, rice, sorghum, rye, millet, etc.Oil-seed plants include cotton, soybean, safflower, sunflower, Brassica,maize, alfalfa, palm, coconut, flax, castor, olive, etc. Leguminousplants include beans and peas. Beans include guar, locust bean,fenugreek, soybean, garden beans, cowpea, mung bean, lima bean, favabean, lentils, chickpea, etc.

Evaluation of Plant Transformation

Following introduction of heterologous foreign DNA into plant cells, thetransformation or integration of heterologous gene in the plant genomeis confirmed by various methods such as analysis of nucleic acids,proteins and metabolites associated with the integrated gene.

PCR analysis is a rapid method to screen transformed cells, tissue orshoots for the presence of incorporated gene at the earlier stage beforetransplanting into the soil (Sambrook and Russell, (2001) MolecularCloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, ColdSpring Harbor, NY). PCR is carried out using oligonucleotide primersspecific to the gene of interest or Agrobacterium vector background,etc.

Plant transformation may be confirmed by Southern blot analysis ofgenomic DNA (Sambrook and Russell, (2001) supra). In general, total DNAis extracted from the transformant, digested with appropriaterestriction enzymes, fractionated in an agarose gel and transferred to anitrocellulose or nylon membrane. The membrane or “blot” is then probedwith, for example, radiolabeled 32P target DNA fragment to confirm theintegration of introduced gene into the plant genome according tostandard techniques (Sambrook and Russell, (2001) supra).

In Northern blot analysis, RNA is isolated from specific tissues oftransformant, fractionated in a formaldehyde agarose gel, and blottedonto a nylon filter according to standard procedures that are routinelyused in the art (Sambrook and Russell, (2001) supra). Expression of RNAencoded by the pesticidal gene is then tested by hybridizing the filterto a radioactive probe derived from a pesticidal gene (Sambrook andRussell, (2001) supra).

Western blot, biochemical assays and the like may be carried out on thetransgenic plants to confirm the presence of protein encoded by thepesticidal gene by standard procedures (Sambrook and Russell, 2001,supra) using antibodies that bind to one or more epitopes present on theIPD113 polypeptide.

Methods to Introduce Genome Editing Technologies into Plants

In some embodiments, the disclosed IPD113 polynucleotide compositionscan be introduced into the genome of a plant using genome editingtechnologies, or previously introduced IPD113 polynucleotides in thegenome of a plant may be edited using genome editing technologies. Forexample, the disclosed polynucleotides can be introduced into a desiredlocation in the genome of a plant through the use of double-strandedbreak technologies such as TALENs, meganucleases, zinc finger nucleases,CRISPR-Cas, and the like. For example, the disclosed polynucleotides canbe introduced into a desired location in a genome using a CRISPR-Cassystem, for the purpose of site-specific insertion. The desired locationin a plant genome can be any desired target site for insertion, such asa genomic region amenable for breeding or may be a target site locatedin a genomic window with an existing trait of interest. Existing traitsof interest could be either an endogenous trait or a previouslyintroduced trait.

In some embodiments, where the disclosed IPD113 polynucleotide haspreviously been introduced into a genome, genome editing technologiesmay be used to alter or modify the introduced polynucleotide sequence.Site specific modifications that can be introduced into the disclosedIPD113 polynucleotide compositions include those produced using anymethod for introducing site specific modification, including, but notlimited to, through the use of gene repair oligonucleotides (e.g. USPublication 2013/0019349), or through the use of double-stranded breaktechnologies such as TALENs, meganucleases, zinc finger nucleases,CRISPR-Cas, and the like. Such technologies can be used to modify thepreviously introduced polynucleotide through the insertion, deletion orsubstitution of nucleotides within the introduced polynucleotide.Alternatively, double-stranded break technologies can be used to addadditional nucleotide sequences to the introduced polynucleotide.Additional sequences that may be added include, additional expressionelements, such as enhancer and promoter sequences. In anotherembodiment, genome editing technologies may be used to positionadditional insecticidally-active proteins in close proximity to thedisclosed IPD113 polynucleotide compositions disclosed herein within thegenome of a plant, to generate molecular stacks of insecticidally-activeproteins.

An “altered target site,” “altered target sequence.” “modified targetsite,” and “modified target sequence” are used interchangeably hereinand refer to a target sequence as disclosed herein that comprises atleast one alteration when compared to non-altered target sequence. Such“alterations” include, for example: (i) replacement of at least onenucleotide, (ii) a deletion of at least one nucleotide, (iii) aninsertion of at least one nucleotide, or (iv) any combination of(i)-(iii).

Stacking of Traits in Transgenic Plant

Transgenic plants may comprise a stack of one or more insecticidalpolynucleotides disclosed herein with one or more additionalpolynucleotides resulting in the production or suppression of multiplepolypeptide sequences. Transgenic plants comprising stacks ofpolynucleotide sequences can be obtained by either or both oftraditional breeding methods or through genetic engineering methods.These methods include, but are not limited to, breeding individual lineseach comprising a polynucleotide of interest, transforming a transgenicplant comprising a gene disclosed herein with a subsequent gene andco-transformation of genes into a single plant cell. As used herein, theterm “stacked” includes having the multiple traits present in the sameplant (i.e., both traits are incorporated into the nuclear genome, onetrait is incorporated into the nuclear genome and one trait isincorporated into the genome of a plastid or both traits areincorporated into the genome of a plastid). In one non-limiting example,“stacked traits” comprise a molecular stack where the sequences arephysically adjacent to each other. A trait, as used herein, refers tothe phenotype derived from a particular sequence or groups of sequences.Co-transformation of genes can be carried out using singletransformation vectors comprising multiple genes or genes carriedseparately on multiple vectors. If the sequences are stacked bygenetically transforming the plants, the polynucleotide sequences ofinterest can be combined at any time and in any order. The traits can beintroduced simultaneously in a co-transformation protocol with thepolynucleotides of interest provided by any combination oftransformation cassettes. For example, if two sequences will beintroduced, the two sequences can be contained in separatetransformation cassettes (trans) or contained on the same transformationcassette (cis). Expression of the sequences can be driven by the samepromoter or by different promoters. In certain cases, it may bedesirable to introduce a transformation cassette that will suppress theexpression of the polynucleotide of interest. This may be combined withany combination of other suppression cassettes or overexpressioncassettes to generate the desired combination of traits in the plant. Itis further recognized that polynucleotide sequences can be stacked at adesired genomic location using a site-specific recombination system.See, for example, WO 1999/25821, WO 1999/25854, WO 1999/25840, WO1999/25855 and WO 1999/25853, all of which are herein incorporated byreference.

In some embodiments, the polynucleotides encoding the IPD113 polypeptidedisclosed herein, alone or stacked with one or more additional insectresistance traits can be stacked with one or more additional inputtraits (e.g., herbicide resistance, fungal resistance, virus resistance,stress tolerance, disease resistance, male sterility, stalk strength,and the like) or output traits (e.g., increased yield, modifiedstarches, improved oil profile, balanced amino acids, high lysine ormethionine, increased digestibility, improved fiber quality, droughtresistance, and the like). Thus, the polynucleotide embodiments can beused to provide a complete agronomic package of improved crop qualitywith the ability to flexibly and cost effectively control any number ofagronomic pests.

Transgenes useful for stacking include but are not limited to:

1. Transgenes that Confer Resistance to Insects or Disease and thatEncode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example, Jones, et al., (1994) Science266:789 (cloning of the tomato Cf-9 gene for resistance to Cladosporiumfulvum); Martin, et al., (1993) Science 262:1432 (tomato Pto gene forresistance to Pseudomonas syringae pv. tomato encodes a protein kinase);Mindrinos, et al., (1994) Cell 78:1089 (Arabidopsis RSP2 gene forresistance to Pseudomonas syringae), McDowell and Woffenden, (2003)Trends Biotechnol. 21(4):178-83 and Toyoda, et al., (2002) TransgenicRes. 11(6):567-82. A plant resistant to a disease is one that is moreresistant to a pathogen as compared to the wild type plant.

(B) Genes encoding a Bacillus thuringiensis protein, a derivativethereof or a synthetic polypeptide modeled thereon. See, for example,Geiser, et al., (1986) Gene 48:109, who disclose the cloning andnucleotide sequence of a Bt delta-endotoxin gene. Moreover, DNAmolecules encoding delta-endotoxin genes can be purchased from AmericanType Culture Collection (Rockville, Md.), for example, under ATCC®Accession Numbers 40098, 67136, 31995 and 31998. Other non-limitingexamples of Bacillus thuringiensis transgenes being geneticallyengineered are given in the following patents and patent applicationsand hereby are incorporated by reference for this purpose: U.S. Pat.Nos. 5,188,960; 5,689,052; 5,880,275; 5,986,177; 6,023,013, 6,060,594,6,063,597, 6,077,824, 6,620,988, 6,642,030, 6,713,259, 6,893,826,7,105,332; 7,179,965, 7,208,474; 7,227,056, 7,288,643, 7,323,556,7,329,736, 7,449,552, 7,468,278, 7,510,878, 7,521,235, 7,544,862,7,605,304, 7,696,412, 7,629,504, 7,705,216, 7,772,465, 7,790,846,7,858,849 and WO 1991/14778; WO 1999/31248; WO 2001/12731; WO 1999/24581and WO 1997/40162.

Genes encoding pesticidal proteins may also be stacked including, butare not limited to: insecticidal proteins from Pseudomonas sp. such asPSEEN3174 (Monalysin; (2011) PLoS Pathogens 7:1-13); from Pseudomonasprotegens strain CHAO and Pf-5 (previously fluorescens) (Pechy-Tarr,(2008) Environmental Microbiology 10:2368-2386; GenBank Accession No.EU400157); from Pseudomonas taiwanensis (Liu, et al., (2010) J. Agric.Food

Chem., 58:12343-12349) and from Pseudomonas pseudoalcaligenes (Zhang, etal., (2009) Annals of Microbiology 59:45-50 and Li, et al., (2007) PlantCell Tiss. Organ Cult. 89:159-168); insecticidal proteins fromPhotorhabdus sp. and Xenorhabdus sp. (Hinchliffe, et al., (2010) TheOpen Toxicology Journal, 3:101-118 and Morgan, et al., (2001) Appliedand Envir. Micro. 67:2062-2069); U.S. Pat. Nos. 6,048,838, and6,379,946; a PIP-1 polypeptide of U.S. Pat. No. 9,688,730; an AfIP-1Aand/or AfIP-1B polypeptide of U.S. Pat. No. 9,475,847; a PIP-47polypeptide of US Publication Number US20160186204; an IPD045polypeptide, an IPD064 polypeptide, an IPD074 polypeptide, an IPD075polypeptide, and an IPD077 polypeptide of PCT Publication Number WO2016/114973; an IPD080 polypeptide of PCT Serial Number PCT/US17/56517;an IPD078 polypeptide, an IPD084 polypeptide, an IPD085 polypeptide, anIPD086 polypeptide, an IPD087 polypeptide, an IPD088 polypeptide, and anIPD089 polypeptide of Serial Number PCT/US17/54160; PIP-72 polypeptideof US Patent Publication Number US20160366891; a PtIP-50 polypeptide anda PtIP-65 polypeptide of US Publication Number US20170166921; an IPD098polypeptide, an IPD059 polypeptide, an IPD108 polypeptide, an IPD109polypeptide of US Serial number 62/521084; a PtIP-83 polypeptide of USPublication Number US20160347799; a PtIP-96 polypeptide of USPublication Number US20170233440; an IPD079 polypeptide of PCTPublication Number WO2017/23486; an IPD082 polypeptide of PCTPublication Number WO 2017/105987, an IPD090 polypeptide of SerialNumber PCT/US17/30602, an IPD093 polypeptide of U.S. Ser. No. 62/434020;an IPD103 polypeptide of Serial Number PCT/US17/39376; an IPD101polypeptide of U.S. Ser. No. 62/438179; an IPD121 polypeptide of U.S.Ser. No. 62/508,514; and δ-endotoxins including, but not limited to aCry1, Cry2, Cry3, Cry4, Cry5, Cry6, Cry7, Cry8, Cry9, Cry10, Cry11,Cry12, Cry13, Cry14, Cry15, Cry16, Cry17, Cry18, Cry19, Cry20, Cry21,Cry22, Cry23, Cry24, Cry25, Cry26, Cry27, Cry28, Cry29, Cry30, Cry31,Cry32, Cry33, Cry34, Cry35,Cry36, Cry37, Cry38, Cry39, Cry40, Cry41,Cry42, Cry43, Cry44, Cry45, Cry46, Cry47, Cry49, Cry50, Cry51, Cry52,Cry53, Cry54, Cry55, Cry56, Cry57, Cry58, Cry59, Cry60, Cry61, Cry62,Cry63, Cry64, Cry65, Cry66, Cry67, Cry68, Cry69, Cry70, Cry71, and Cry72 classes of δ-endotoxin polypeptides and the B. thuringiensiscytolytic cyt1 and cyt2 genes. Members of these classes of B.thuringiensis insecticidal proteins (see, Crickmore, et al., “Bacillusthuringiensis toxin nomenclature” (2011), and can be found atlifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/, which can be accessed onthe world-wide web using the “www” prefix). Examples of δ-endotoxinsalso include but are not limited to Cry1A proteins of U.S. Pat. Nos.5,880,275, 7,858,849, and 8,878,007; a Cry1Ac mutant of U.S. Pat. No.9,512,187; a DIG-3 or DIG-11 toxin (N-terminal deletion of α-helix 1and/or α-helix 2 variants of cry proteins such as Cry1A, Cry3A) of U.S.Pat. Nos. 8,304,604, 8.304,605 and 8,476,226; Cry1B of U.S. patentapplication Ser. No. 10/525,318, US Patent Application PublicationNumber US20160194364, and U.S. Pat. Nos. 9,404,121 and 8,772,577; Cry1Bvariants of PCT Publication Number WO2016/61197 and Serial NumberPCT/US17/27160; Cry1C of U.S. Pat. No. 6,033,874; Cry1D protein ofUS20170233759; a Cry1E protein of PCT Serial Number PCT/US17/53178; aCry1F protein of U.S. Pat. Nos. 5,188,960 and 6,218,188; Cry1A/Fchimeras of U.S. Pat. Nos. 7,070,982; 6,962,705 and 6,713,063; a Cry1Iprotein of PCT Publication number WO 2017/0233759; a Cry1J variant of USPublication US20170240603; a Cry2 protein such as Cry2Ab protein of U.S.Pat. No. 7,064,249 and Cry2A.127 protein of U.S. Pat. No. 7,208,474; aCry3A protein including but not limited to an engineered hybridinsecticidal protein (eHIP) created by fusing unique combinations ofvariable regions and conserved blocks of at least two different Cryproteins (US Patent Application Publication Number 2010/0017914); a Cry4protein; a Cry5 protein; a Cry6 protein; Cry8 proteins of U.S. Pat. Nos.7,329,736, 7,449,552, 7,803,943, 7,476,781, 7,105,332, 7,339,092,7,378,499, 7,462,760, and 9,593,345; a Cry9 protein such as such asmembers of the Cry9A, Cry9B, Cry9C, Cry9D, Cry9E and Cry9F familiesincluding the Cry9 protein of U.S. Pat. Nos. 9,000,261 and 8,802,933,and US Serial Number WO 2017/132188; a Cry15 protein of Naimov, et al.,(2008) Applied and Environmental Microbiology, 74:7145-7151; a Cry14protein of U.S. Pat. No. US8,933,299; a Cry22, a Cry34Ab1 protein ofU.S. Pat. Nos. 6,127,180, 6,624,145 and 6,340,593; a truncated Cry34protein of U.S. Pat. No. 8,816,157; a CryET33 and cryET34 protein ofU.S. Pat. Nos. 6,248,535, 6,326,351, 6,399,330, 6,949,626, 7,385,107 and7,504,229; a CryET33 and CryET34 homologs of US Patent PublicationNumber 2006/0191034, 2012/0278954, and PCT Publication Number WO2012/139004; a Cry35Ab1 protein of U.S. Pat. Nos. 6,083,499, 6,548,291and 6,340,593; a Cry46 protein of U.S. Pat. No. 9,403,881, a Cry 51protein, a Cry binary toxin; a TIC901 or related toxin; TIC807 of USPatent Application Publication Number 2008/0295207; TIC853 of U.S. Pat.No. 8,513,493; ET29, ET37, TIC809, TIC810, TIC812, TIC127, TIC128 of PCTUS 2006/033867; engineered Hemipteran toxic proteins of US PatentApplication Publication Number US20160150795, AXMI-027, AXMI-036, andAXMI-038 of U.S. Pat. No. 8,236,757; AXMI-031, AXMI-039, AXMI-040,AXMI-049 of U.S. Pat. No. 7,923,602; AXMI-018, AXMI-020 and AXMI-021 ofWO 2006/083891; AXMI-010 of WO 2005/038032; AXMI-003 of WO 2005/021585;AXMI-008 of US Patent Application Publication Number 2004/0250311;AXMI-006 of US Patent Application Publication Number 2004/0216186;AXMI-007 of US Patent Application Publication Number 2004/0210965;AXMI-009 of US Patent Application Number 2004/0210964; AXMI-014 of USPatent Application Publication Number 2004/0197917; AXMI-004 of USPatent Application Publication Number 2004/0197916; AXMI-028 andAXMI-029 of WO 2006/119457; AXMI-007, AXMI-008, AXMI-0080rf2, AXMI-009,AXMI-014 and AXMI-004 of WO 2004/074462; AXMI-150 of U.S. Pat. No.8,084,416; AXMI-205 of US Patent Application Publication Number2011/0023184; AXMI-011, AXMI-012, AXMI-013, AXMI-015, AXMI-019,AXMI-044, AXMI-037, AXMI-043, AXMI-033, AXMI-034, AXMI-022, AXMI-023,AXMI-041, AXMI-063 and AXMI-064 of US Patent Application PublicationNumber 2011/0263488; AXMI046, AXMI048, AXMI050, AXMI051, AXMI052,AXMI053, AXMI054, AXMI055, AXMI056, AXMI057, AXMI058, AXMI059, AXMI060,AXMI061, AXMI067, AXMI069, AXMI071, AXMI072, AXMI073, AXMI074, AXMI075,AXMI087, AXMI088, AXMI093, AXMI070, AXMI080, AXMI081, AXMI082, AXMI091,AXMI092, AXMI096, AXMI097, AXMI098, AXMI099, AXMI100, AXMI101, AXMI102,AXMI103, AXMI104, AXMI107, AXMI108, AXMI109, AXMI110, AXMI111, AXMI112,AXMI114, AXMI116, AXMI117, AXMI118, AXMI119, AXMI120, AXMI121, AXMI122,AXMI123, AXMI124, AXMI125, AXMI126, AXMI127, AXMI129, AXMI151, AXMI161,AXMI164, AXMI183, AXMI132, AXMI137, AXMI138 of U.S. Pat. Nos. 8461421and 8,461,422; AXMI-R1 and related proteins of US Patent ApplicationPublication Number 2010/0197592; AXMI221Z, AXMI222z, AXMI223z, AXMI224zand AXMI225z of WO 2011/103248; AXMI218, AXMI219, AXMI220, AXMI226,AXMI227, AXMI228, AXMI229, AXMI230 and AXMI231 of WO 2011/103247;AXMI-115, AXMI-113, AXMI-005, AXMI-163 and AXMI-184 of U.S. Pat. No.8,334,431; AXMI-001, AXMI-002, AXMI-030, AXMI-035 and AXMI-045 of USPatent Application Publication Number 2010/0298211; AXMI-066 andAXMI-076 of US Patent Application Publication Number 2009/0144852;AXMI128, AXMI130, AXMI131, AXMI133, AXMI140, AXMI141, AXMI142, AXMI143,AXMI144, AXMI146, AXMI148, AXMI149, AXMI152, AXMI153, AXMI154, AXMI155,AXMI156, AXMI157, AXMI158, AXMI162, AXMI165, AXMI166, AXMI167, AXMI168,AXMI169, AXMI170, AXMI171, AXMI172, AXMI173, AXMI174, AXMI175, AXMI176,AXMI177, AXMI178, AXMI179, AXMI180, AXMI181, AXMI182, AXMI185, AXMI186,AXMI187, AXMI188, AXMI189 of U.S. Pat. No. 8,318,900; AXMI079, AXMI080,AXMI081, AXMI082, AXMI091, AXMI092, AXMI096, AXMI097, AXMI098, AXMI099,AXMI100, AXMI101, AXMI102, AXMI103, AXMI104, AXMI107, AXMI108, AXMI109,AXMI110, dsAXMI111, AXMI112, AXMI114, AXMI116, AXMI117, AXMI118,AXMI119, AXMI120, AXMI121, AXMI122, AXMI123, AXMI124, AXMI1257,AXMI1268, AXMI127, AXMI129, AXMI164, AXMI151, AXMI161, AXMI183, AXMI132,AXMI138, AXMI137 of US Patent US8461421; AXMI192 of US PatentUS8,461,415; AXMI281 of US Patent Application Publication NumberUS20160177332; AXMI422 of US Pat. No. US8,252,872; cry proteins such asCry1A and Cry3A having modified proteolytic sites of U.S. Pat. No.8,319,019; a Cry1Ac, Cry2Aa and Cry1Ca toxin protein from Bacillusthuringiensis strain VBTS 2528 of US Patent Application PublicationNumber 2011/0064710. The Cry proteins MP032, MP049, MP051, MP066, MP068,MP070, MP091S, MP109S, MP114, MP121, MP134S, MP183S, MP185S, MP186S,MP195S, MP197S, MP208S, MP209S, MP212S, MP214S, MP217S, MP222S, MP234S,MP235S, MP237S, MP242S, MP243, MP248, MP249S, MP251 M, MP252S, MP253,MP259S, MP287S, MP288S, MP295S, MP296S, MP297S, MP300S, MP304S, MP306S,MP310S, MP312S, MP314S, MP319S, MP325S, MP326S, MP327S, MP328S, MP334S,MP337S, MP342S, MP349S, MP356S, MP359S, MP360S, MP437S, MP451S, MP452S,MP466S, MP468S, MP476S, MP482S, MP522S, MP529S, MP548S, MP552S, MP562S,MP564S, MP566S, MP567S, MP569S, MP573S, MP574S, MP575S, MP581S, MP590,MP594S, MP596S, MP597, MP599S, MP600S, MP601S, MP602S, MP604S, MP626S,MP629S, MP630S, MP631S, MP632S, MP633S, MP634S, MP635S, MP639S, MP640S,MP644S, MP649S, MP651S, MP652S, MP653S, MP661S, MP666S, MP672S, MP696S,MP704S, MP724S, MP729S, MP739S, MP755S, MP773S, MP799S, MP800S, MP801S,MP802S, MP803S, MP805S, MP809S, MP815S, MP828S, MP831S, MP844S, MP852,MP865S, MP879S, MP887S, MP891S, MP896S, MP898S, MP935S, MP968, MP989,MP993, MP997, MP1049, MP1066, MP1067, MP1080, MP1081, MP1200, MP1206,MP1233, and MP1311 of US Serial Number 62/607372. The insecticidalactivity of Cry proteins is well known to one skilled in the art (forreview, see, van Frannkenhuyzen, (2009) J. Invert. Path. 101:1-16). Theuse of Cry proteins as transgenic plant traits is well known to oneskilled in the art and Cry-transgenic plants including but not limitedto plants expressing Cry1Ac, Cry1Ac+Cry2Ab, Cry1Ab, Cry1A.105, Cry1F,Cry1Fa2, Cry1F+Cry1Ac, Cry2Ab, Cry3A, mCry3A, Cry3Bb1, Cry34Ab1,Cry35Ab1, Vip3A, mCry3A, Cry9c and CBI-Bt have received regulatoryapproval (see, Sanahuja, (2011) Plant Biotech Journal 9:283-300 and theCERA. (2010) GM Crop Database Center for Environmental Risk Assessment(CERA), ILSI Research Foundation, Washington D.C. atcera-gmc.org/index.php?action=gm_crop_database which can be accessed onthe world-wide web using the “www” prefix). More than one pesticidalproteins can also be expressed in plants such as Vip3Ab & Cry1Fa(US2012/0317682); Cry1BE & Cry1F (US2012/0311746); Cry1CA & Cry1AB(US2012/0311745); Cry1F & CryCa (US2012/0317681); Cry1DA & Cry1BE(US2012/0331590); Cry1DA & Cry1Fa (US2012/0331589); Cry1AB & Cry1BE(US2012/0324606); Cry1Fa & Cry2Aa and Cry1I & Cry1E (US2012/0324605);Cry34Ab/35Ab & Cry6Aa (US20130167269); Cry34Ab/VCry35Ab & Cry3Aa(US20130167268); Cry1Da & Cry1Ca (U.S. Pat. No. 9,796,982); Cry3Aa &Cry6Aa (U.S. Pat. No. 9,798,963); and Cry3A & Cry1Ab or Vip3Aa (U.S.Pat. No. 9,045,766). Pesticidal proteins also include insecticidallipases including lipid acyl hydrolases of U.S. Pat. No. 7,491,869, andcholesterol oxidases such as from Streptomyces (Purcell et al. (1993)Biochem Biophys Res Commun 15:1406-1413). Pesticidal proteins alsoinclude VIP (vegetative insecticidal proteins) toxins of U.S. Pat. Nos.5,877,012, 6,107,279 6,137,033, 7,244,820, 7,615,686, and 8,237,020 andthe like. Other VIP proteins are well known (see,Iifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/vip.html which can beaccessed on the world-wide web using the “www” prefix). Pesticidalproteins also include Cyt proteins including Cyt1A variants of PCTSerial Number PCT/US2017/000510; Pesticidal proteins also include toxincomplex (TC) proteins, obtainable from organisms such as Xenorhabdus,Photorhabdus and Paenibacillus (see, U.S. Pat. Nos. 7,491,698 and8,084,418). Some TC proteins have “stand alone” insecticidal activityand other TC proteins enhance the activity of the stand-alone toxinsproduced by the same given organism. The toxicity of a “stand-alone” TCprotein (from Photorhabdus, Xenorhabdus or Paenibacillus, for example)can be enhanced by one or more TC protein “potentiators” derived from asource organism of a different genus. There are three main types of TCproteins. As referred to herein, Class A proteins (“Protein A”) arestand-alone toxins. Class B proteins (“Protein B”) and Class C proteins(“Protein C”) enhance the toxicity of Class A proteins. Examples ofClass A proteins are TcbA, TcdA, XptA1 and XptA2. Examples of Class Bproteins are TcaC, TcdB, XptB1Xb and XptC1Wi. Examples of Class Cproteins are TccC, XptC1Xb and XptB1Wi. Pesticidal proteins also includespider, snake and scorpion venom proteins. Examples of spider venompeptides include but not limited to lycotoxin-1 peptides and mutantsthereof (U.S. Pat. No. 8,334,366).

(C) A polynucleotide encoding an insect-specific hormone or pheromonesuch as an ecdysteroid and juvenile hormone, a variant thereof, amimetic based thereon or an antagonist or agonist thereof. See, forexample, the disclosure by Hammock, et al., (1990) Nature 344:458, ofbaculovirus expression of cloned juvenile hormone esterase, aninactivator of juvenile hormone.

(D) A polynucleotide encoding an insect-specific peptide which, uponexpression, disrupts the physiology of the affected pest. For example,see the disclosures of, Regan, (1994) J. Biol. Chem. 269:9 (expressioncloning yields DNA coding for insect diuretic hormone receptor); Pratt,et al., (1989) Biochem. Biophys. Res. Comm. 163:1243 (an allostatin isidentified in Diploptera puntata); Chattopadhyay, et al., (2004)Critical Reviews in Microbiology 30(1):33-54; Zjawiony, (2004) J NatProd 67(2):300-310; Carlini and Grossi-de-Sa, (2002) Toxicon40(11):1515-1539; Ussuf, et al., (2001) Curr Sci. 80(7):847-853 andVasconcelos and Oliveira, (2004) Toxicon 44(4):385-403. See also, U.S.Pat. No. 5,266,317 to Tomalski, et al., who disclose genes encodinginsect-specific toxins.

(E) A polynucleotide encoding an enzyme responsible for ahyperaccumulation of a monoterpene, a sesquiterpene, a steroid,hydroxamic acid, a phenylpropanoid derivative or another non-proteinmolecule with insecticidal activity.

(F) A polynucleotide encoding an enzyme involved in the modification,including the post-translational modification, of a biologically activemolecule; for example, a glycolytic enzyme, a proteolytic enzyme, alipolytic enzyme, a nuclease, a cyclase, a transaminase, an esterase, ahydrolase, a phosphatase, a kinase, a phosphorylase, a polymerase, anelastase, a chitinase and a glucanase, whether natural or synthetic.See, PCT Application WO 1993/02197 in the name of Scott, et al., whichdiscloses the nucleotide sequence of a callase gene. DNA molecules whichcontain chitinase-encoding sequences can be obtained, for example, fromthe ATCC® under Accession Numbers 39637 and 67152. See also, Kramer, etal., (1993) Insect Biochem. Molec. Biol. 23:691, who teach thenucleotide sequence of a cDNA encoding tobacco hookworm chitinase andKawalleck, et al., (1993) Plant Molec. Biol. 21:673, who provide thenucleotide sequence of the parsley ubi4-2 polyubiquitin gene, and U.S.Pat. Nos. 6,563,020; 7,145,060 and 7,087,810.

(G) A polynucleotide encoding a molecule that stimulates signaltransduction. For example, see the disclosure by Botella, et al., (1994)Plant Molec. Biol. 24:757, of nucleotide sequences for mung beancalmodulin cDNA clones, and Griess, et al., (1994) Plant Physiol.104:1467, who provide the nucleotide sequence of a maize calmodulin cDNAclone.

(H) A polynucleotide encoding a hydrophobic moment peptide. See, PCTApplication WO 1995/16776 and U.S. Pat. No. 5,580,852 disclosure ofpeptide derivatives of Tachyplesin which inhibit fungal plant pathogens)and PCT Application WO 1995/18855 and U.S. Pat. No. 5,607,914 (teachessynthetic antimicrobial peptides that confer disease resistance).

(I) A polynucleotide encoding a membrane permease, a channel former or achannel blocker. For example, see the disclosure by Jaynes, et al.,(1993) Plant Sci. 89:43, of heterologous expression of a cecropin-betalytic peptide analog to render transgenic tobacco plants resistant toPseudomonas solanacearum.

(J) A gene encoding a viral-invasive protein or a complex toxin derivedtherefrom. For example, the accumulation of viral coat proteins intransformed plant cells imparts resistance to viral infection and/ordisease development effected by the virus from which the coat proteingene is derived, as well as by related viruses. See, Beachy, et al.,(1990) Ann. Rev. Phytopathol. 28:451. Coat protein-mediated resistancehas been conferred upon transformed plants against alfalfa mosaic virus,cucumber mosaic virus, tobacco streak virus, potato virus X, potatovirus Y, tobacco etch virus, tobacco rattle virus and tobacco mosaicvirus. Id.

(K) A gene encoding an insect-specific antibody or an immunotoxinderived therefrom. Thus, an antibody targeted to a critical metabolicfunction in the insect gut would inactivate an affected enzyme, killingthe insect. Cf. Taylor, et al., Abstract #497, SEVENTH INT'L SYMPOSIUMON MOLECULAR PLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994)(enzymatic inactivation in transgenic tobacco via production ofsingle-chain antibody fragments).

(L) A gene encoding a virus-specific antibody. See, for example,Tavladoraki, et al., (1993) Nature 366:469, who show that transgenicplants expressing recombinant antibody genes are protected from virusattack.

(M) A polynucleotide encoding a developmental-arrestive protein producedin nature by a pathogen or a parasite. Thus, fungal endoalpha-1,4-D-polygalacturonases facilitate fungal colonization and plantnutrient release by solubilizing plant cell wallhomo-alpha-1,4-D-galacturonase. See, Lamb, et al., (1992) Bio/Technology10:1436. The cloning and characterization of a gene which encodes a beanendopolygalacturonase-inhibiting protein is described by Toubart, etal., (1992) Plant J. 2:367.

(N) A polynucleotide encoding a developmental-arrestive protein producedin nature by a plant. For example, Logemann, et al., (1992)Bio/Technology 10:305, have shown that transgenic plants expressing thebarley ribosome-inactivating gene have an increased resistance to fungaldisease.

(O) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, (1995) Current Biology5(2), Pieterse and Van Loon, (2004) Curr. Opin. Plant Bio. 7(4):456-64and Somssich, (2003) Cell 113(7):815-6.

(P) Antifungal genes (Cornelissen and Melchers, (1993) Pl. Physiol.101:709-712 and Parijs, et al., (1991) Planta 183:258-264 and Bushnell,et al., (1998) Can. J. of Plant Path. 20(2):137-149. Also see, U.S.patent application Ser. Nos. 09/950,933; 11/619,645; 11/657,710;11/748,994; 11/774,121 and U.S. Pat. Nos. 6,891,085 and 7,306,946. LysMReceptor-like kinases for the perception of chitin fragments as a firststep in plant defense response against fungal pathogens (US2012/0110696).

(Q) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see, U.S. Pat. Nos. 5,716,820; 5,792,931; 5,798,255;5,846,812; 6,083,736; 6,538,177; 6,388,171 and 6,812,380.

(R) A polynucleotide encoding a Cystatin and cysteine proteinaseinhibitors. See, U.S. Pat. No. 7,205,453.

(S) Defensin genes. See, WO 2003/000863 and U.S. Pat. Nos. 6,911,577;6,855,865; 6,777,592 and 7,238,781.

(T) Genes conferring resistance to nematodes. See, e.g., PCT ApplicationWO 1996/30517; PCT Application WO 1993/19181, WO 2003/033651 and Urwin,et al., (1998) Planta 204:472-479, Williamson, (1999) Curr Opin PlantBio. 2(4):327-31; U.S. Pat. Nos. 6,284,948 and 7,301,069 and miR164genes (WO 2012/058266).

(U) Genes that confer resistance to Phytophthora Root Rot, such as theRps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes.See, for example, Shoemaker, et al., Phytophthora Root Rot ResistanceGene Mapping in Soybean, Plant Genome IV Conference, San Diego, Calif.(1995).

(V) Genes that confer resistance to Brown Stem Rot, such as described inU.S. Pat. No. 5,689,035 and incorporated by reference for this purpose.

(W) Genes that confer resistance to Colletotrichum, such as described inUS Patent Application Publication US 2009/0035765 and incorporated byreference for this purpose. This includes the Rcg locus that may beutilized as a single locus conversion.

2. Transgenes that Confer Resistance to a Herbicide, for Example:

(A) A polynucleotide encoding resistance to a herbicide that inhibitsthe growing point or meristem, such as an imidazolinone or asulfonylurea. Exemplary genes in this category code for mutant ALS andAHAS enzyme as described, for example, by Lee, et al., (1988) EMBO J.7:1241 and Miki, et al., (1990) Theor. Appl. Genet. 80:449,respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659; 5,141,870;5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107; 5,928,937 and5,378,824; U.S. patent application Ser. No. 11/683,737 and InternationalPublication WO 1996/33270.

(B) A polynucleotide encoding a protein for resistance to Glyphosate(resistance imparted by mutant 5-enolpyruvl-3-phosphikimate synthase(EPSP) and aroA genes, respectively) and other phosphono compounds suchas glufosinate (phosphinothricin acetyl transferase (PAT) andStreptomyces hygroscopicus phosphinothricin acetyl transferase (bar)genes), and pyridinoxy or phenoxy proprionic acids and cyclohexones(ACCase inhibitor-encoding genes). See, for example, U.S. Pat. No.4,940,835 to Shah, et al., which discloses the nucleotide sequence of aform of EPSPS which can confer glyphosate resistance. U.S. Pat. No.5,627,061 to Barry, et al., also describes genes encoding EPSPS enzymes.See also, U.S. Pat. Nos. 6,566,587; 6,338,961; 6,248,876; 6,040,497;5,804,425; 5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642;5,094,945, 4,940,835; 5,866,775; 6,225,114; 6,130,366; 5,310,667;4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449; RE 37,287 E and5,491,288 and International Publications EP 1173580; WO 2001/66704; EP1173581 and EP 1173582, which are incorporated herein by reference forthis purpose. Glyphosate resistance is also imparted to plants thatexpress a gene encoding a glyphosate oxido-reductase enzyme as describedmore fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, which areincorporated herein by reference for this purpose. In additionglyphosate resistance can be imparted to plants by the over expressionof genes encoding glyphosate N-acetyltransferase. See, for example, U.S.Pat. Nos. 7,462,481; 7,405,074 and US Patent Application PublicationNumber US 2008/0234130. A DNA molecule encoding a mutant aroA gene canbe obtained under ATCC® Accession Number 39256, and the nucleotidesequence of the mutant gene is disclosed in U.S. Pat. No. 4,769,061 toComai. EP Application Number 0 333 033 to Kumada, et al., and U.S. Pat.No. 4,975,374 to Goodman, et al., disclose nucleotide sequences ofglutamine synthetase genes which confer resistance to herbicides such asL-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in EP ApplicationNumbers 0 242 246 and 0 242 236 to Leemans, et al.,; De Greef, et al.,(1989) Bio/Technology 7:61, describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. See also, U.S. Pat. Nos. 5,969,213; 5,489,520;5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024;6,177,616 and 5,879,903, which are incorporated herein by reference forthis purpose. Exemplary genes conferring resistance to phenoxyproprionic acids and cyclohexones, such as sethoxydim and haloxyfop, arethe Acc1-S1, Acc1-S2 and Acc1-S3 genes described by Marshall, et al.,(1992) Theor. Appl. Genet. 83:435.

(C) A polynucleotide encoding a protein for resistance to herbicide thatinhibits photosynthesis, such as a triazine (psbA and gs+genes) and abenzonitrile (nitrilase gene). Przibilla, et al., (1991) Plant Cell3:169, describe the transformation of Chlamydomonas with plasmidsencoding mutant psbA genes. Nucleotide sequences for nitrilase genes aredisclosed in U.S. Pat. No. 4,810,648 to Stalker and DNA moleculescontaining these genes are available under ATCC® Accession Numbers53435, 67441 and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes, et al., (1992) Biochem.J. 285:173.

(D) A polynucleotide encoding a protein for resistance to Acetohydroxyacid synthase, which has been found to make plants that express thisenzyme resistant to multiple types of herbicides, has been introducedinto a variety of plants (see, e.g., Hattori, et al., (1995) Mol GenGenet. 246:419). Other genes that confer resistance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota, et al., (1994)Plant Physiol 106:17), genes for glutathione reductase and superoxidedismutase (Aono, et al., (1995) Plant Cell Physiol 36:1687) and genesfor various phosphotransferases (Datta, et al., (1992) Plant Mol Biol20:619).

(E) A polynucleotide encoding resistance to a herbicide targetingProtoporphyrinogen oxidase (protox) which is necessary for theproduction of chlorophyll. The protox enzyme serves as the target for avariety of herbicidal compounds. These herbicides also inhibit growth ofall the different species of plants present, causing their totaldestruction. The development of plants containing altered protoxactivity which are resistant to these herbicides are described in U.S.Pat. Nos. 6,288,306; 6,282,83 and 5,767,373 and InternationalPublication WO 2001/12825.

(F) The aad-1 gene (originally from Sphingobium herbicidovorans) encodesthe aryloxyalkanoate dioxygenase (AAD-1) protein. The trait conferstolerance to 2,4-dichlorophenoxyacetic acid and aryloxyphenoxypropionate(commonly referred to as “fop” herbicides such as quizalofop)herbicides. The aad-1 gene, itself, for herbicide tolerance in plantswas first disclosed in WO 2005/107437 (see also, US 2009/0093366). Theaad-12 gene, derived from Delftia acidovorans, which encodes thearyloxyalkanoate dioxygenase (AAD-12) protein that confers tolerance to2,4-dichlorophenoxyacetic acid and pyridyloxyacetate herbicides bydeactivating several herbicides with an aryloxyalkanoate moiety,including phenoxy auxin (e.g., 2,4-D, MCPA), as well as pyridyloxyauxins (e.g., fluroxypyr, triclopyr).

(G) A polynucleotide encoding a herbicide resistant dicambamonooxygenase disclosed in US Patent Application Publication2003/0135879 for imparting dicamba tolerance;

(H) A polynucleotide molecule encoding bromoxynil nitrilase (Bxn)disclosed in U.S. Pat. No. 4,810,648 for imparting bromoxynil tolerance;

(I) A polynucleotide molecule encoding phytoene (crtl) described inMisawa, et al., (1993) Plant J. 4:833-840 and in Misawa, et al., (1994)Plant J. 6:481-489 for norflurazon tolerance.

3. Transgenes that Confer or Contribute to an Altered GrainCharacteristic

-   Such as:

(A) Altered fatty acids, for example, by

(1) Down-regulation of stearoyl-ACP to increase stearic acid content ofthe plant. See, Knultzon, et al., (1992) Proc. Natl. Acad. Sci. USA89:2624 and WO 1999/64579 (Genes to Alter Lipid Profiles in Corn).

(2) Elevating oleic acid via FAD-2 gene modification and/or decreasinglinolenic acid via FAD-3 gene modification (see, U.S. Pat. Nos.6,063,947; 6,323,392; 6,372,965 and WO 1993/11245).

(3) Altering conjugated linolenic or linoleic acid content, such as inWO 2001/12800.

(4) Altering LEC1, AGP, Dek1, Superal1, mi1 ps, and various Ipa genessuch as Ipa1, Ipa3, hpt or hggt. For example, see, WO 2002/42424, WO1998/22604, WO 2003/011015, WO 2002/057439, WO 2003/011015, U.S. Pat.Nos. 6,423,886, 6,197,561, 6,825,397 and US Patent ApplicationPublication Numbers US 2003/0079247, US 2003/0204870 and Rivera-Madrid,et al., (1995) Proc. Natl. Acad. Sci. 92:5620-5624.

(5) Genes encoding delta-8 desaturase for making long-chainpolyunsaturated fatty acids (U.S. Pat. Nos. 8,058,571 and 8,338,152),delta-9 desaturase for lowering saturated fats (U.S. Pat. No.8,063,269), Primula Δ6-desaturase for improving omega-3 fatty acidprofiles.

(6) Isolated nucleic acids and proteins associated with lipid and sugarmetabolism regulation, in particular, lipid metabolism protein (LMP)used in methods of producing transgenic plants and modulating levels ofseed storage compounds including lipids, fatty acids, starches or seedstorage proteins and use in methods of modulating the seed size, seednumber, seed weights, root length and leaf size of plants (EP 2404499).

(7) Altering expression of a High-Level Expression of Sugar-Inducible 2(HSI2) protein in the plant to increase or decrease expression of HSI2in the plant. Increasing expression of HSI2 increases oil content whiledecreasing expression of HSI2 decreases abscisic acid sensitivity and/orincreases drought resistance (US Patent Application Publication Number2012/0066794).

(8) Expression of cytochrome b5 (Cb5) alone or with FAD2 to modulate oilcontent in plant seed, particularly to increase the levels of omega-3fatty acids and improve the ratio of omega-6 to omega-3 fatty acids (USPatent Application Publication Number 2011/0191904).

(9) Nucleic acid molecules encoding wrinkled1-like polypeptides formodulating sugar metabolism (U.S. Pat. No. 8,217,223).

(B) Altered phosphorus content, for example, by the

(1) Introduction of a phytase-encoding gene would enhance breakdown ofphytate, adding more free phosphate to the transformed plant. Forexample, see, Van Hartingsveldt, et al., (1993) Gene 127:87, for adisclosure of the nucleotide sequence of an Aspergillus niger phytasegene.

(2) Modulating a gene that reduces phytate content. In maize, this, forexample, could be accomplished, by cloning and then re-introducing DNAassociated with one or more of the alleles, such as the LPA alleles,identified in maize mutants characterized by low levels of phytic acid,such as in WO 2005/113778 and/or by altering inositol kinase activity asin WO 2002/059324, US Patent Application Publication Number2003/0009011, WO 2003/027243, US Patent Application Publication Number2003/0079247, WO 1999/05298, U.S. Pat. Nos. 6,197,561, 6,291,224,6,391,348, WO 2002/059324, US Patent Application Publication Number2003/0079247, WO 1998/45448, WO 1999/55882, WO 2001/04147.

(C) Altered carbohydrates affected, for example, by altering a gene foran enzyme that affects the branching pattern of starch or, a genealtering thioredoxin such as NTR and/or TRX (see, U.S. Pat. No.6,531,648. which is incorporated by reference for this purpose) and/or agamma zein knock out or mutant such as cs27 or TUSC27 or en27 (see, U.S.Pat. No. 6,858,778 and US Patent Application Publication Number2005/0160488, US Patent Application Publication Number 2005/0204418,which are incorporated by reference for this purpose). See, Shiroza, etal., (1988) J. Bacteriol. 170:810 (nucleotide sequence of Streptococcusmutant fructosyltransferase gene), Steinmetz, et al., (1985) Mol. Gen.Genet. 200:220 (nucleotide sequence of Bacillus subtilis levansucrasegene), Pen, et al., (1992) Bio/Technology 10:292 (production oftransgenic plants that express Bacillus licheniformis alpha-amylase),Elliot, et al., (1993) Plant Molec. Biol. 21:515 (nucleotide sequencesof tomato invertase genes), Søgaard, et al., (1993) J. Biol. Chem.268:22480 (site-directed mutagenesis of barley alpha-amylase gene) andFisher, et al., (1993) Plant Physiol. 102:1045 (maize endosperm starchbranching enzyme II), WO 1999/10498 (improved digestibility and/orstarch extraction through modification of UDP-D-xylose 4-epimerase,Fragile 1 and 2, Ref1, HCHL, C4H), U.S. Pat. No. 6,232,529 (method ofproducing high oil seed by modification of starch levels (AGP)). Thefatty acid modification genes mentioned herein may also be used toaffect starch content and/or composition through the interrelationshipof the starch and oil pathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see, U.S. Pat. No. 6,787,683,US Patent Application Publication Number 2004/0034886 and WO 2000/68393involving the manipulation of antioxidant levels and WO 2003/082899through alteration of a homogentisate geranyl geranyl transferase(hggt).

(E) Altered essential seed amino acids. For example, see, U.S. Pat. No.6,127,600 (method of increasing accumulation of essential amino acids inseeds), U.S. Pat. No. 6,080,913 (binary methods of increasingaccumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389(high lysine), WO 1999/40209 (alteration of amino acid compositions inseeds), WO 1999/29882 (methods for altering amino acid content ofproteins), U.S. Pat. No. 5,850,016 (alteration of amino acidcompositions in seeds), WO 1998/20133 (proteins with enhanced levels ofessential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S.Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plantamino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increasedlysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophansynthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolicenzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414(increased methionine), WO 1998/56935 (plant amino acid biosyntheticenzymes), WO 1998/45458 (engineered seed protein having higherpercentage of essential amino acids), WO 1998/42831 (increased lysine),U.S. Pat. No. 5,633,436 (increasing sulfur amino acid content), U.S.Pat. No. 5,559,223 (synthetic storage proteins with defined structurecontaining programmable levels of essential amino acids for improvementof the nutritional value of plants), WO 1996/01905 (increasedthreonine), WO 1995/15392 (increased lysine), US Patent ApplicationPublication Number 2003/0163838, US Patent Application PublicationNumber 2003/0150014, US Patent Application Publication Number2004/0068767, U.S. Pat. No. 6,803,498, WO 2001/79516.

4. Genes that Control Male-Sterility:

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar, et al., and chromosomal translocationsas described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Inaddition to these methods, Albertsen, et al., U.S. Pat. No. 5,432,068,describe a system of nuclear male sterility which includes: identifyinga gene which is critical to male fertility; silencing this native genewhich is critical to male fertility; removing the native promoter fromthe essential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing or turning “on”,the promoter, which in turn allows the gene that, confers male fertilityto be transcribed.

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT (WO 2001/29237).

(B) Introduction of various stamen-specific promoters (WO 1992/13956, WO1992/13957).

(C) Introduction of the barnase and the barstar gene (Paul, et al.,(1992) Plant Mol. Biol. 19:611-622).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426; 5,478,369;5,824,524; 5,850,014 and 6,265,640, all of which are hereby incorporatedby reference.

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.For example, see, Lyznik, et al., (2003) Plant Cell Rep 21:925-932 andWO 1999/25821, which are hereby incorporated by reference. Other systemsthat may be used include the Gin recombinase of phage Mu (Maeser, etal., (1991) Vicki Chandler, The Maize Handbook ch. 118 (Springer-Verlag1994), the Pin recombinase of E. coli (Enomoto, et al., 1983) and theR/RS system of the pSRi plasmid (Araki, et al., 1992).

6. Genes that Affect Abiotic Stress Resistance

Including but not limited to flowering, ear and seed development,enhancement of nitrogen utilization efficiency, altered nitrogenresponsiveness, drought resistance or tolerance, cold resistance ortolerance and salt resistance or tolerance and increased yield understress.

(A) For example, see: WO 2000/73475 where water use efficiency isaltered through alteration of malate; U.S. Pat. Nos. 5,892,009,5,965,705, 5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866,6,717,034, 6,801,104, WO 2000/060089, WO 2001/026459, WO 2001/035725, WO2001/034726, WO 2001/035727, WO 2001/036444, WO 2001/036597, WO2001/036598, WO 2002/015675, WO 2002/017430, WO 2002/077185, WO2002/079403, WO 2003/013227, WO 2003/013228, WO 2003/014327, WO2004/031349, WO 2004/076638, WO 199809521.

(B) WO 199938977 describing genes, including CBF genes and transcriptionfactors effective in mitigating the negative effects of freezing, highsalinity and drought on plants, as well as conferring other positiveeffects on plant phenotype.

(C) US Patent Application Publication Number 2004/0148654 and WO2001/36596 where abscisic acid is altered in plants resulting inimproved plant phenotype such as increased yield and/or increasedtolerance to abiotic stress.

(D) WO 2000/006341, WO 2004/090143, U.S. Pat. Nos. 7,531,723 and6,992,237 where cytokinin expression is modified resulting in plantswith increased stress tolerance, such as drought tolerance, and/orincreased yield. Also see, WO 2002/02776, WO 2003/052063, JP2002/281975, U.S. Pat. No. 6,084,153, WO 2001/64898, U.S. Pat. Nos.6,177,275 and 6,107,547 (enhancement of nitrogen utilization and alterednitrogen responsiveness).

(E) For ethylene alteration, see, US Patent Application PublicationNumber 2004/0128719, US Patent Application Publication Number2003/0166197 and WO 2000/32761.

(F) For plant transcription factors or transcriptional regulators ofabiotic stress, see, e.g., US Patent Application Publication Number2004/0098764 or US Patent Application Publication Number 2004/0078852.

(G) Genes that increase expression of vacuolar pyrophosphatase such asAVP1 (U.S. Pat. No. 8,058,515) for increased yield; nucleic acidencoding a HSFA4 or a HSFAS (Heat Shock Factor of the class A4 or A5)polypeptides, an oligopeptide transporter protein (OPT4-like)polypeptide; a plastochron2-like (PLA2-like) polypeptide or a Wuschelrelated homeobox 1-like (WOX1-like) polypeptide (U. Patent ApplicationPublication Number US 2011/0283420).

(H) Down regulation of polynucleotides encoding poly (ADP-ribose)polymerase (PARP) proteins to modulate programmed cell death (U.S. Pat.No. 8,058,510) for increased vigor.

(I) Polynucleotide encoding DTP21 polypeptides for conferring droughtresistance (US Patent Application Publication Number US 2011/0277181).

(J) Nucleotide sequences encoding ACC Synthase 3 (ACS3) proteins formodulating development, modulating response to stress, and modulatingstress tolerance (US Patent Application Publication Number US2010/0287669).

(K) Polynucleotides that encode proteins that confer a drought tolerancephenotype (DTP) for conferring drought resistance (WO 2012/058528).

(L) Tocopherol cyclase (TC) genes for conferring drought and salttolerance (US Patent Application Publication Number 2012/0272352).

(M) CAAX amino terminal family proteins for stress tolerance (U.S. Pat.No. 8,338,661).

(N) Mutations in the SAL1 encoding gene have increased stress tolerance,including increased drought resistant (US Patent Application PublicationNumber 2010/0257633).

(O) Expression of a nucleic acid sequence encoding a polypeptideselected from the group consisting of: GRF polypeptide, RAA1-likepolypeptide, SYR polypeptide, ARKL polypeptide, and YTP polypeptideincreasing yield-related traits (US Patent Application PublicationNumber 2011/0061133).

(P) Modulating expression in a plant of a nucleic acid encoding a ClassIII Trehalose Phosphate Phosphatase (TPP) polypeptide for enhancingyield-related traits in plants, particularly increasing seed yield (USPatent Application Publication Number 2010/0024067).

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see e.g., WO1997/49811 (LHY), WO 1998/56918 (ESD4), WO 1997/10339 and U.S. Pat. No.6,573,430 (TFL), U.S. Pat. No. 6,713,663 (FT), WO 1 996/1 441 4 (CON),WO 1996/38560, WO 2001/21822 (VRN1), WO 2000/44918 (VRN2), WO 1999/49064(GI), WO 2000/46358 (FR1), WO 1997/29123, U.S. Pat. Nos. 6,794,560,6,307,126 (GAI), WO 1999/09174 (D8 and Rht) and WO 2004/076638 and WO2004/031349 (transcription factors).

7. Genes that Confer Increased Yield

(A) A transgenic crop plant transformed by a1-AminoCyclopropane-1-Carboxylate Deaminase-like Polypeptide (ACCDP)coding nucleic acid, wherein expression of the nucleic acid sequence inthe crop plant results in the plant's increased root growth, and/orincreased yield, and/or increased tolerance to environmental stress ascompared to a wild type variety of the plant (U.S. Pat. No. 8,097,769).

(B) Over-expression of maize zinc finger protein gene (Zm-ZFP1) using aseed preferred promoter has been shown to enhance plant growth, increasekernel number and total kernel weight per plant (US Patent ApplicationPublication Number 2012/0079623).

(C) Constitutive over-expression of maize lateral organ boundaries (LOB)domain protein (Zm-LOBDP1) has been shown to increase kernel number andtotal kernel weight per plant (US Patent Application Publication Number2012/0079622).

(D) Enhancing yield-related traits in plants by modulating expression ina plant of a nucleic acid encoding a VIM1 (Variant in Methylation1)-like polypeptide or a VTC2-like (GDP-L-galactose phosphorylase)polypeptide or a DUF1685 polypeptide or an ARF6-like (Auxin ResponsiveFactor) polypeptide (WO 2012/038893).

(E) Modulating expression in a plant of a nucleic acid encoding aSte20-like polypeptide or a homologue thereof gives plants havingincreased yield relative to control plants (EP 2431472).

(F) Genes encoding nucleoside diphosphatase kinase (NDK) polypeptidesand homologs thereof for modifying the plant's root architecture (USPatent Application Publication Number 2009/0064373).

8. Genes that confer plant digestibility.

(A) Altering the level of xylan present in the cell wall of a plant bymodulating expression of xylan synthase (U.S. Pat. No. 8,173,866).

In some embodiments, the stacked trait may be a trait or event that hasreceived regulatory approval including but not limited to the eventswith regulatory and can be found at the Center for Environmental RiskAssessment (cera-gmc.org/?action=gm_crop_database, which can be accessedusing the www prefix) and at the International Service for theAcquisition of Agri-Biotech Applications(isaaa.org/gmapprovaldatabase/default.asp, which can be accessed usingthe www prefix).

Gene Silencing

In some embodiments, the stacked trait may be in the form of silencingof one or more polynucleotides of interest resulting in suppression ofone or more target pest polypeptides. In some embodiments, the silencingis achieved using a suppression DNA construct.

In some embodiments, one or more polynucleotide encoding thepolypeptides of the IPD113 polypeptide or fragments or variants thereofmay be stacked with one or more polynucleotides encoding one or morepolypeptides having insecticidal activity or agronomic traits as setforth supra and optionally may further include one or morepolynucleotides providing for gene silencing of one or more targetpolynucleotides as discussed infra.

“Suppression DNA construct” is a recombinant DNA construct which whentransformed or stably integrated into the genome of the plant, resultsin “silencing” of a target gene in the plant. The target gene may beendogenous or transgenic to the plant. “Silencing,” as used herein withrespect to the target gene, refers generally to the suppression oflevels of mRNA or protein/enzyme expressed by the target gene, and/orthe level of the enzyme activity or protein functionality. The term“suppression” includes lower, reduce, decline, decrease, inhibit,eliminate and prevent. “Silencing” or “gene silencing” does not specifymechanism and is inclusive, and not limited to, anti-sense,cosuppression, viral-suppression, hairpin suppression, stem-loopsuppression, RNAi-based approaches and small RNA-based approaches.

A suppression DNA construct may comprise a region derived from a targetgene of interest and may comprise all or part of the nucleic acidsequence of the sense strand (or antisense strand) of the target gene ofinterest. Depending upon the approach to be utilized, the region may be100% identical or less than 100% identical (e.g., at least 50% or anyinteger between 51% and 100% identical) to all or part of the sensestrand (or antisense strand) of the gene of interest.

Suppression DNA constructs are readily constructed once the target geneof interest is selected, and include, without limitation, cosuppressionconstructs, antisense constructs, viral-suppression constructs, hairpinsuppression constructs, stem-loop suppression constructs,double-stranded RNA-producing constructs, and more generally, RNAi (RNAinterference) constructs and small RNA constructs such as siRNA (shortinterfering RNA) constructs and miRNA (microRNA) constructs.

“Antisense inhibition” refers to the production of antisense RNAtranscripts capable of suppressing the expression of the target protein.“Antisense RNA” refers to an RNA transcript that is complementary to allor part of a target primary transcript or mRNA and that blocks theexpression of a target isolated nucleic acid fragment. Thecomplementarity of an antisense RNA may be with any part of the specificgene transcript, i.e., at the 5′ non-coding sequence, 3′ non-codingsequence, introns or the coding sequence.

“Cosuppression” refers to the production of sense RNA transcriptscapable of suppressing the expression of the target protein. “Sense” RNArefers to RNA transcript that includes the mRNA and can be translatedinto protein within a cell or in vitro. Cosuppression constructs inplants have been previously designed by focusing on overexpression of anucleic acid sequence having homology to a native mRNA, in the senseorientation, which results in the reduction of all RNA having homologyto the overexpressed sequence (see, Vaucheret, et al., (1998) Plant J.16:651-659 and Gura, (2000) Nature 404:804-808).

Another variation describes the use of plant viral sequences to directthe suppression of proximal mRNA encoding sequences (PCT Publication WO1998/36083).

Recent work has described the use of “hairpin” structures thatincorporate all or part, of an mRNA encoding sequence in a complementaryorientation that results in a potential “stem-loop” structure for theexpressed RNA (PCT Publication WO 1999/53050). In this case the stem isformed by polynucleotides corresponding to the gene of interest insertedin either sense or anti-sense orientation with respect to the promoterand the loop is formed by some polynucleotides of the gene of interest,which do not have a complement in the construct. This increases thefrequency of cosuppression or silencing in the recovered transgenicplants. For review of hairpin suppression, see, Wesley, et al., (2003)Methods in Molecular Biology, Plant Functional Genomics: Methods andProtocols 236:273-286.

A construct where the stem is formed by at least 30 nucleotides from agene to be suppressed and the loop is formed by a random nucleotidesequence has also effectively been used for suppression (PCT PublicationWO 1999/61632).

The use of poly-T and poly-A sequences to generate the stem in thestem-loop structure has also been described (PCT Publication WO2002/00894).

Yet another variation includes using synthetic repeats to promoteformation of a stem in the stem-loop structure. Transgenic organismsprepared with such recombinant DNA fragments have been shown to havereduced levels of the protein encoded by the nucleotide fragment formingthe loop as described in PCT Publication WO 2002/00904.

RNA interference refers to the process of sequence-specificpost-transcriptional gene silencing in animals mediated by shortinterfering RNAs (siRNAs) (Fire, et al., (1998) Nature 391:806). Thecorresponding process in plants is commonly referred to aspost-transcriptional gene silencing (PTGS) or RNA silencing and is alsoreferred to as quelling in fungi. The process of post-transcriptionalgene silencing is thought to be an evolutionarily-conserved cellulardefense mechanism used to prevent the expression of foreign genes and iscommonly shared by diverse flora and phyla (Fire, et al., (1999) TrendsGenet. 15:358). Such protection from foreign gene expression may haveevolved in response to the production of double-stranded RNAs (dsRNAs)derived from viral infection or from the random integration oftransposon elements into a host genome via a cellular response thatspecifically destroys homologous single-stranded RNA of viral genomicRNA. The presence of dsRNA in cells triggers the RNAi response through amechanism that has yet to be fully characterized.

The presence of long dsRNAs in cells stimulates the activity of aribonuclease III enzyme referred to as dicer. Dicer is involved in theprocessing of the dsRNA into short pieces of dsRNA known as shortinterfering RNAs (siRNAs) (Berstein, et al., (2001) Nature 409:363).Short interfering RNAs derived from dicer activity are typically about21 to about 23 nucleotides in length and comprise about 19 base pairduplexes (Elbashir, et al., (2001) Genes Dev. 15:188). Dicer has alsobeen implicated in the excision of 21- and 22-nucleotide small temporalRNAs (stRNAs) from precursor RNA of conserved structure that areimplicated in translational control (Hutvagner, et al., (2001) Science293:834). The RNAi response also features an endonuclease complex,commonly referred to as an RNA-induced silencing complex (RISC), whichmediates cleavage of single-stranded RNA having sequence complementarityto the antisense strand of the siRNA duplex. Cleavage of the target RNAtakes place in the middle of the region complementary to the antisensestrand of the siRNA duplex (Elbashir, et al., (2001) Genes Dev. 15:188).In addition, RNA interference can also involve small RNA (e.g., miRNA)mediated gene silencing, presumably through cellular mechanisms thatregulate chromatin structure and thereby prevent transcription of targetgene sequences (see, e.g., Allshire, (2002) Science 297:1818-1819;Volpe, et al., (2002) Science 297:1833-1837; Jenuwein, (2002) Science297:2215-2218 and Hall, et al., (2002) Science 297:2232-2237). As such,miRNA molecules of the disclosure can be used to mediate gene silencingvia interaction with RNA transcripts or alternately by interaction withparticular gene sequences, wherein such interaction results in genesilencing either at the transcriptional or post-transcriptional level.

Methods and compositions are further provided which allow for anincrease in RNAi produced from the silencing element. In suchembodiments, the methods and compositions employ a first polynucleotidecomprising a silencing element for a target pest sequence operablylinked to a promoter active in the plant cell; and, a secondpolynucleotide comprising a suppressor enhancer element comprising thetarget pest sequence or an active variant or fragment thereof operablylinked to a promoter active in the plant cell. The combined expressionof the silencing element with suppressor enhancer element leads to anincreased amplification of the inhibitory RNA produced from thesilencing element over that achievable with only the expression of thesilencing element alone. In addition to the increased amplification ofthe specific RNAi species itself, the methods and compositions furtherallow for the production of a diverse population of RNAi species thatcan enhance the effectiveness of disrupting target gene expression. Assuch, when the suppressor enhancer element is expressed in a plant cellin combination with the silencing element, the methods and compositioncan allow for the systemic production of RNAi throughout the plant; theproduction of greater amounts of RNAi than would be observed with justthe silencing element construct alone; and, the improved loading of RNAiinto the phloem of the plant, thus providing better control of phloemfeeding insects by an RNAi approach. Thus, the various methods andcompositions provide improved methods for the delivery of inhibitory RNAto the target organism. See, for example, US Patent ApplicationPublication 2009/0188008.

As used herein, a “suppressor enhancer element” comprises apolynucleotide comprising the target sequence to be suppressed or anactive fragment or variant thereof. It is recognized that the suppressorenhancer element need not be identical to the target sequence, butrather, the suppressor enhancer element can comprise a variant of thetarget sequence, so long as the suppressor enhancer element hassufficient sequence identity to the target sequence to allow for anincreased level of the RNAi produced by the silencing element over thatachievable with only the expression of the silencing element. Similarly,the suppressor enhancer element can comprise a fragment of the targetsequence, wherein the fragment is of sufficient length to allow for anincreased level of the RNAi produced by the silencing element over thatachievable with only the expression of the silencing element.

It is recognized that multiple suppressor enhancer elements from thesame target sequence or from different target sequences or fromdifferent regions of the same target sequence can be employed. Forexample, the suppressor enhancer elements employed can comprisefragments of the target sequence derived from different region of thetarget sequence (i.e., from the 3′UTR, coding sequence, intron, and/or5′UTR). Further, the suppressor enhancer element can be contained in anexpression cassette, as described elsewhere herein, and in specificembodiments, the suppressor enhancer element is on the same or on adifferent

DNA vector or construct as the silencing element. The suppressorenhancer element can be operably linked to a promoter as disclosedherein. It is recognized that the suppressor enhancer element can beexpressed constitutively or alternatively, it may be produced in astage-specific manner employing the various inducible ortissue-preferred or developmentally regulated promoters that arediscussed elsewhere herein.

In specific embodiments, employing both a silencing element and thesuppressor enhancer element the systemic production of RNAi occursthroughout the entire plant. In further embodiments, the plant or plantparts of the disclosure have an improved loading of RNAi into the phloemof the plant than would be observed with the expression of the silencingelement construct alone and, thus provide better control of phloemfeeding insects by an RNAi approach. In specific embodiments, theplants, plant parts and plant cells of the disclosure can further becharacterized as allowing for the production of a diversity of RNAispecies that can enhance the effectiveness of disrupting target geneexpression.

In specific embodiments, the combined expression of the silencingelement and the suppressor enhancer element increases the concentrationof the inhibitory RNA in the plant cell, plant, plant part, plant tissueor phloem over the level that is achieved when the silencing element isexpressed alone.

As used herein, an “increased level of inhibitory RNA” comprises anystatistically significant increase in the level of RNAi produced in aplant having the combined expression when compared to an appropriatecontrol plant. For example, an increase in the level of RNAi in theplant, plant part or the plant cell can comprise at least about a 1%,about a 1%-5%, about a 5%-10%, about a 10%-20%, about a 20%-30%, about a30%-40%, about a 40%-50%, about a 50%-60%, about 60-70%, about 70%-80%,about a 80%-90%, about a 90%-100% or greater increase in the level ofRNAi in the plant, plant part, plant cell or phloem when compared to anappropriate control. In other embodiments, the increase in the level ofRNAi in the plant, plant part, plant cell or phloem can comprise atleast about a 1 fold, about a 1 fold-5 fold, about a 5 fold-10 fold,about a 10 fold-20 fold, about a 20 fold-30 fold, about a 30 fold-40fold, about a 40 fold-50 fold, about a 50 fold-60 fold, about 60 fold-70fold, about 70 fold-80 fold, about a 80 fold-90 fold, about a 90fold-100 fold or greater increase in the level of RNAi in the plant,plant part, plant cell or phloem when compared to an appropriatecontrol. Examples of combined expression of the silencing element withsuppressor enhancer element for the control of Stinkbugs and Lygus canbe found in US Patent Application Publication 2011/0301223 and US PatentApplication Publication 2009/0192117.

Some embodiments relate to down-regulation of expression of target genesin insect pest species by interfering ribonucleic acid (RNA) molecules.PCT Publication WO 2007/074405 describes methods of inhibitingexpression of target genes in invertebrate pests including Coloradopotato beetle. PCT Publication WO 2005/110068 describes methods ofinhibiting expression of target genes in invertebrate pests includingWestern corn rootworm to control insect infestation. Furthermore, PCTPublication WO 2009/091864 describes compositions and methods for thesuppression of target genes from insect pest species including pestsfrom the Lygus genus. Nucleic acid molecules including RNAi fortargeting the vacuolar ATPase H subunit, useful for controlling acoleopteran pest population and infestation as described in US PatentApplication Publication 2012/0198586. PCT Publication WO 2012/055982describes ribonucleic acid (RNA or double stranded RNA) that inhibits ordown regulates the expression of a target gene that encodes: an insectribosomal protein such as the ribosomal protein L19, the ribosomalprotein L40 or the ribosomal protein S27A; an insect proteasome subunitsuch as the Rpn6 protein, the Pros 25, the Rpn2 protein, the proteasomebeta 1 subunit protein or the Pros beta 2 protein; an insect β-coatomerof the COPI vesicle, the γ-coatomer of the COPI vesicle, the β′-coatomerprotein or the ζ-coatomer of the COPI vesicle; an insect Tetraspanine 2A protein which is a putative transmembrane domain protein; an insectprotein belonging to the actin family such as Actin 5C; an insectubiquitin-5E protein; an insect Sec23 protein which is a GTPaseactivator involved in intracellular protein transport; an insectcrinkled protein which is an unconventional myosin which is involved inmotor activity; an insect crooked neck protein which is involved in theregulation of nuclear alternative mRNA splicing; an insect vacuolarH+-ATPase G-subunit protein and an insect Tbp-1 such as Tat-bindingprotein. PCT publication WO 2007/035650 describes ribonucleic acid (RNAor double stranded RNA) that inhibits or down regulates the expressionof a target gene that encodes Snf7. US Patent Application publication2011/0054007 describes polynucleotide silencing elements targetingRPS10. US Patent Application publications 2014/0275208 andUS2015/0257389 describe polynucleotide silencing elements targetingRyanR and PAT3. PCT Patent Application publication WO2016/138106describes polynucleotide silencing elements targeting coatomer alpha orgamma. US Patent Application Publications 2012/029750, US 20120297501,and 2012/0322660 describe interfering ribonucleic acids (RNA or doublestranded RNA) that functions upon uptake by an insect pest species todown-regulate expression of a target gene in said insect pest, whereinthe RNA comprises at least one silencing element wherein the silencingelement is a region of double-stranded RNA comprising annealedcomplementary strands, one strand of which comprises or consists of asequence of nucleotides which is at least partially complementary to atarget nucleotide sequence within the target gene. US Patent ApplicationPublication 2012/0164205 describe potential targets for interferingdouble stranded ribonucleic acids for inhibiting invertebrate pestsincluding: a Chd3 Homologous Sequence, a Beta-Tubulin HomologousSequence, a 40 kDa V-ATPase Homologous Sequence, a EF1a HomologousSequence, a 26S Proteosome Subunit p28 Homologous Sequence, a JuvenileHormone Epoxide Hydrolase Homologous Sequence, a Swelling DependentChloride Channel Protein Homologous Sequence, a Glucose-6-Phosphate1-Dehydrogenase Protein Homologous Sequence, an Act42A ProteinHomologous Sequence, a ADP-Ribosylation Factor 1 Homologous Sequence, aTranscription Factor IIB Protein Homologous Sequence, a ChitinaseHomologous Sequences, a Ubiquitin Conjugating Enzyme HomologousSequence, a Glyceraldehyde-3-Phosphate Dehydrogenase HomologousSequence, an Ubiquitin B Homologous Sequence, a Juvenile HormoneEsterase Homolog, and an Alpha Tubuliln Homologous Sequence.

Use in Pesticidal Control

General methods for employing strains comprising a nucleic acid sequenceof the embodiments or a variant thereof, in pesticide control or inengineering other organisms as pesticidal agents are known in the art.

Microorganism hosts that are known to occupy the “phytosphere”(phylloplane, phyllosphere, rhizosphere, and/or rhizoplana) of one ormore crops of interest may be selected. These microorganisms areselected to be capable of successfully competing in the particularenvironment with the wild-type microorganisms, provide for stablemaintenance and expression of the gene expressing the IPD113 polypeptideand desirably provide for improved protection of the pesticide fromenvironmental degradation and inactivation.

Alternatively, the IPD113 polypeptide is produced by introducing aheterologous gene into a cellular host. Expression of the heterologousgene results, directly or indirectly, in the intracellular productionand maintenance of the pesticide. These cells are then treated underconditions that prolong the activity of the toxin produced in the cellwhen the cell is applied to the environment of target pest(s). Theresulting product retains the toxicity of the toxin. These naturallyencapsulated IPD113 polypeptides may then be formulated in accordancewith conventional techniques for application to the environment hostinga target pest, e.g., soil, water, and foliage of plants. See, forexample EPA 0192319, and the references cited therein.

Pesticidal Compositions

In some embodiments, the active ingredients can be applied in the formof compositions and can be applied to the crop area or plant to betreated, simultaneously or in succession, with other compounds. Thesecompounds can be fertilizers, weed killers, Cryoprotectants,surfactants, detergents, pesticidal soaps, dormant oils, polymers,and/or time-release or biodegradable carrier formulations that permitlong-term dosing of a target area following a single application of theformulation. They can also be selective herbicides, chemicalinsecticides, virucides, microbicides, amoebicides, pesticides,fungicides, bacteriocides, nematocides, molluscicides or mixtures ofseveral of these preparations, if desired, together with furtheragriculturally acceptable carriers, surfactants or application-promotingadjuvants customarily employed in the art of formulation. Suitablecarriers and adjuvants can be solid or liquid and correspond to thesubstances ordinarily employed in formulation technology, e.g. naturalor regenerated mineral substances, solvents, dispersants, wettingagents, tackifiers, binders or fertilizers. Likewise, the formulationsmay be prepared into edible “baits” or fashioned into pest “traps” topermit feeding or ingestion by a target pest of the pesticidalformulation.

Methods of applying an active ingredient or an agrochemical compositionthat contains at least one of the IPD113 polypeptide produced by thebacterial strains include leaf application, seed coating and soilapplication. The number of applications and the rate of applicationdepend on the intensity of infestation by the corresponding pest.

The composition may be formulated as a powder, dust, pellet, granule,spray, emulsion, colloid, solution or such like, and may be prepared bysuch conventional means as desiccation, lyophilization, homogenation,extraction, filtration, centrifugation, sedimentation or concentrationof a culture of cells comprising the polypeptide. In all suchcompositions that contain at least one such pesticidal polypeptide, thepolypeptide may be present in a concentration of from about 1% to about99% by weight.

Lepidopteran, Dipteran, Heteropteran, nematode, Hemiptera or Coleopteranpests may be killed or reduced in numbers in each area by the methods ofthe disclosure or may be prophylactically applied to an environmentalarea to prevent infestation by a susceptible pest. Preferably the pestingests or is contacted with, a pesticidally-effective amount of thepolypeptide. “Pesticidally-effective amount” as used herein refers to anamount of the pesticide that can bring about death to at least one pestor to noticeably reduce pest growth, feeding or normal physiologicaldevelopment. This amount will vary depending on such factors as, forexample, the specific target pests to be controlled, the specificenvironment, location, plant, crop or agricultural site to be treated,the environmental conditions and the method, rate, concentration,stability, and quantity of application of the pesticidally-effectivepolypeptide composition. The formulations may also vary with respect toclimatic conditions, environmental considerations, and/or frequency ofapplication and/or severity of pest infestation.

The pesticide compositions described may be made by formulating thebacterial cell, Crystal and/or spore suspension or isolated proteincomponent with the desired agriculturally-acceptable carrier. Thecompositions may be formulated prior to administration in an appropriatemeans such as lyophilized, freeze-dried, desiccated or in an aqueouscarrier, medium or suitable diluent, such as saline or another buffer.The formulated compositions may be in the form of a dust or granularmaterial or a suspension in oil (vegetable or mineral) or water oroil/water emulsions or as a wettable powder or in combination with anyother carrier material suitable for agricultural application. Suitableagricultural carriers can be solid or liquid. The term“agriculturally-acceptable carrier” covers all adjuvants, inertcomponents, dispersants, surfactants, tackifiers, binders, etc. that areordinarily used in pesticide formulation technology. The formulationsmay be mixed with one or more solid or liquid adjuvants and prepared byvarious means, e.g., by homogeneously mixing, blending and/or grindingthe pesticidal composition with suitable adjuvants using conventionalformulation techniques. Suitable formulations and application methodsare described in U.S. Pat. No. 6,468,523, herein incorporated byreference. The plants can also be treated with one or more chemicalcompositions, including one or more herbicide, insecticides orfungicides. Exemplary chemical compositions include: Fruits/VegetablesHerbicides: Atrazine, Bromacil, Diuron, Glyphosate, Linuron, Metribuzin,Simazine, Trifluralin, Fluazifop, Glufosinate, Halo sulfuron Gowan,Paraquat, Propyzamide, Sethoxydim, Butafenacil, Halosulfuron,Indaziflam; Fruits/Vegetables Insecticides: Aldicarb, Bacillusthuriengiensis, Carbaryl, Carbofuran, Chlorpyrifos, Cypermethrin,Deltamethrin, Diazinon, Malathion, Abamectin,Cyfluthrin/beta-cyfluthrin, Esfenvalerate, Lambda-cyhalothrin,Acequinocyl, Bifenazate, Methoxyfenozide, Novaluron, Chromafenozide,Thiacloprid, Dinotefuran, FluaCrypyrim, Tolfenpyrad, Clothianidin,Spirodiclofen, Gamma-cyhalothrin, Spiromesifen, Spinosad, Rynaxypyr,Cyazypyr, Spinoteram, Triflumuron, Spirotetramat, Imidacloprid,Flubendiamide, Thiodicarb, Metaflumizone, Sulfoxaflor, Cyflumetofen,Cyanopyrafen, Imidacloprid, Clothianidin, Thiamethoxam, Spinotoram,Thiodicarb, Flonicamid, Methiocarb, Emamectin-benzoate, lndoxacarb,Forthiazate, Fenamiphos, Cadusaphos, Pyriproxifen, Fenbutatin-oxid,Hexthiazox, Methomyl,4-[[(6-Chlorpyridin-3₁1)methyl](2,2-difluorethyl)amino]furan-2(5H)-on;Fruits/Vegetables Fungicides: Carbendazim, Chlorothalonil, EBDCs,Sulphur, Thiophanate-methyl, Azoxystrobin, Cymoxanil, Fluazinam,Fosetyl, Iprodione, Kresoxim-methyl, Metalaxyl/mefenoxam,Trifloxystrobin, Ethaboxam, 1provalicarb, Trifloxystrobin, Fenhexamid,Oxpoconazole fumarate, Cyazofamid, Fenamidone, Zoxamide, Picoxystrobin,Pyraclostrobin, Cyflufenamid, Boscalid; Cereals Herbicides: Isoproturon,Bromoxynil, loxynil, Phenoxies, Chlorsulfuron, Clodinafop, Diclofop,Diflufenican, Fenoxaprop, Florasulam, Fluoroxypyr, Metsulfuron,Triasulfuron, Flucarbazone, lodosulfuron, Propoxycarbazone, Picolinafen,Mesosulfuron, Beflubutamid, Pinoxaden, Amidosulfuron, ThifensulfuronMethyl, Tribenuron, Flupyrsulfuron, Sulfosulfuron, Pyrasulfotole,Pyroxsulam, Flufenacet, Tralkoxydim, Pyroxasulfon; Cereals Fungicides:Carbendazim, Chlorothalonil, Azoxystrobin, Cyproconazole, Cyprodinil,Fenpropimorph, Epoxiconazole, Kresoxim-methyl, Quinoxyfen, Tebuconazole,Trifloxystrobin, Simeconazole, Picoxystrobin, Pyraclostrobin,Dimoxystrobin, Prothioconazole, Fluoxastrobin; Cereals Insecticides:Dimethoate, Lambda-cyhalthrin, Deltamethrin, alpha-Cypermethrin,β-cyfluthrin, Bifenthrin, Imidacloprid, Clothianidin, Thiamethoxam,Thiacloprid, Acetamiprid, Dinetofuran, Clorphyriphos, Metamidophos,Oxidemethon-methyl, Pirimicarb, Methiocarb; Maize Herbicides: Atrazine,Alachlor, Bromoxynil, Acetochlor, Dicamba, Clopyralid, (S-)Dimethenamid, Glufosinate, Glyphosate, Isoxaflutole, (S-)Metolachlor,Mesotrione, Nicosulfuron, Primisulfuron, Rimsulfuron, Sulcotrione,Foramsulfuron, Topramezone, Tembotrione, Saflufenacil, Thiencarbazone,Flufenacet, Pyroxasulfon; Maize Insecticides: Carbofuran, Chlorpyrifos,Bifenthrin, Fipronil, Imidacloprid, Lambda-Cyhalothrin, Tefluthrin,Terbufos, Thiamethoxam, Clothianidin, Spiromesifen, Flubendiamide,Triflumuron, Rynaxypyr, Deltamethrin, Thiodicarb, β-Cyfluthrin,Cypermethrin, Bifenthrin, Lufenuron, Triflumoron,Tefluthrin,Tebupirimphos, Ethiprole, Cyazypyr, Thiacloprid, Acetamiprid,Dinetofuran, Avermectin, Methiocarb, Spirodiclofen, Spirotetramat; MaizeFungicides: Fenitropan, Thiram, Prothioconazole, Tebuconazole,Trifloxystrobin; Rice Herbicides: Butachlor, Propanil, Azimsulfuron,Bensulfuron, Cyhalofop, Daimuron, Fentrazamide, Imazosulfuron,Mefenacet, Oxaziclomefone, Pyrazosulfuron, Pyributicarb, Quinclorac,Thiobencarb, Indanofan, Flufenacet, Fentrazamide, Halosulfuron,Oxaziclomefone, Benzobicyclon, Pyriftalid, Penoxsulam, Bispyribac,Oxadiargyl, Ethoxysulfuron, Pretilachlor, Mesotrione, Tefuryltrione,Oxadiazone, Fenoxaprop, Pyrimisulfan; Rice Insecticides: Diazinon,Fenitrothion, Fenobucarb, Monocrotophos, Benfuracarb, Buprofezin,Dinotefuran, Fipronil, Imidacloprid, Isoprocarb, Thiacloprid,Chromafenozide, Thiacloprid, Dinotefuran, Clothianidin, Ethiprole,Flubendiamide, Rynaxypyr, Deltamethrin, Acetamiprid, Thiamethoxam,Cyazypyr, Spinosad, Spinotoram, Emamectin-Benzoate, Cypermethrin,Chlorpyriphos, Cartap, Methamidophos, Etofenprox, Triazophos,4-[[(6-Chlorpyridin-3-Amethyl](2,2-difluorethyl)amino]furan-2(5H)-on,Carbofuran, Benfuracarb; Rice Fungicides: Thiophanate-methyl,Azoxystrobin, Carpropamid, Edifenphos, Ferimzone, Iprobenfos,Isoprothiolane, Pencycuron, Probenazole, Pyroquilon, Tricyclazole,Trifloxystrobin, Diclocymet, Fenoxanil, Simeconazole, Tiadinil; CottonHerbicides: Diuron, Fluometuron, MSMA, Oxyfluorfen, Prometryn,Trifluralin, Carfentrazone, Clethodim, Fluazifop-butyl, Glyphosate,Norflurazon, Pendimethalin, Pyrithiobac-sodium, Trifloxysulfuron,Tepraloxydim, Glufosinate, Flumioxazin, Thidiazuron; CottonInsecticides: Acephate, Aldicarb, Chlorpyrifos, Cypermethrin,Deltamethrin, Malathion, Monocrotophos, Abamectin, Acetamiprid,Emamectin Benzoate, Imidacloprid, Indoxacarb, Lambda-Cyhalothrin,Spinosad, Thiodicarb, Gamma-Cyhalothrin, Spiromesifen, Pyridalyl,Flonicamid, Flubendiamide, Triflumuron, Rynaxypyr, Beta-Cyfluthrin,Spirotetramat, Clothianidin, Thiamethoxam, Thiacloprid, Dinetofuran,Flubendiamide, Cyazypyr, Spinosad, Spinotoram, gamma Cyhalothrin,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Thiodicarb, Avermectin, Flonicamid, Pyridalyl, Spiromesifen,Sulfoxaflor, Profenophos, Thriazophos, Endosulfan; Cotton Fungicides:Etridiazole, Metalaxyl, Quintozene; Soybean Herbicides: Alachlor,Bentazone, Trifluralin, Chlorimuron-Ethyl, Cloransulam-Methyl,Fenoxaprop, Fomesafen, Fluazifop, Glyphosate, Imazamox, Imazaquin,Imazethapyr, (S-)Metolachlor, Metribuzin, Pendimethalin, Tepraloxydim,Glufosinate; Soybean Insecticides: Lambda-cyhalothrin, Methomyl,Parathion, Thiocarb, Imidacloprid, Clothianidin, Thiamethoxam,Thiacloprid, Acetamiprid, Dinetofuran, Flubendiamide, Rynaxypyr,Cyazypyr, Spinosad, Spinotoram, Emamectin-Benzoate, Fipronil, Ethiprole,Deltamethrin, β-Cyfluthrin, gamma and lambda Cyhalothrin,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Spirotetramat, Spinodiclofen, Triflumuron, Flonicamid, Thiodicarb,beta-Cyfluthrin; Soybean Fungicides: Azoxystrobin, Cyproconazole,Epoxiconazole, Flutriafol, Pyraclostrobin, Tebuconazole,Trifloxystrobin, Prothioconazole, Tetraconazole; Sugarbeet Herbicides:Chloridazon, Desmedipham, Ethofumesate, Phenmedipham, Triallate,Clopyralid, Fluazifop, Lenacil, Metamitron, Quinmerac, Cycloxydim,Triflusulfuron, Tepraloxydim, Quizalofop; Sugarbeet Insecticides:Imidacloprid, Clothianidin, Thiamethoxam, Thiacloprid, Acetamiprid,Dinetofuran, Deltamethrin, β-Cyfluthrin, gamma/lambda Cyhalothrin,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Tefluthrin, Rynaxypyr, Cyaxypyr, Fipronil, Carbofuran; CanolaHerbicides: Clopyralid, Diclofop, Fluazifop, Glufosinate, Glyphosate,Metazachlor, Trifluralin Ethametsulfuron, Quinmerac, Quizalofop,Clethodim, Tepraloxydim; Canola Fungicides: Azoxystrobin, Carbendazim,Fludioxonil, Iprodione, Prochloraz, Vinclozolin; Canola Insecticides:Carbofuran organophosphates, Pyrethroids, Thiacloprid, Deltamethrin,Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dinetofuran,β-Cyfluthrin, gamma and lambda Cyhalothrin, tau-Fluvaleriate, Ethiprole,Spinosad, Spinotoram, Flubendiamide, Rynaxypyr, Cyazypyr,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on.

In some embodiments, the herbicide is Atrazine, Bromacil, Diuron,Chlorsulfuron, Metsulfuron, Thifensulfuron Methyl, Tribenuron,Acetochlor, Dicamba, Isoxaflutole, Nicosulfuron, Rimsulfuron,Pyrithiobac-sodium, Flumioxazin, Chlorimuron-Ethyl, Metribuzin,Quizalofop, S-metolachlor, Hexazinne or combinations thereof.

In some embodiments, the insecticide is Esfenvalerate,Chlorantraniliprole, Methomyl, Indoxacarb, Oxamyl or combinationsthereof.

Pesticidal and insecticidal activity

“Pest” includes but is not limited to, insects, fungi, bacteria,nematodes, mites, ticks and the like. Insect pests include insectsselected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera,Mallophaga, Homoptera, Hemiptera Orthroptera, Thysanoptera, Dermaptera,Isoptera, Anoplura, Siphonaptera, Trichoptera, etc., particularlyLepidoptera and Coleoptera.

Those skilled in the art will recognize that not all compounds areequally effective against all pests. Compounds of the embodimentsdisplay activity against insect pests, which may include economicallyimportant agronomic, forest, greenhouse, nursery ornamentals, food andfiber, public and animal health, domestic and commercial structure,household and stored product pests.

Larvae of the order Lepidoptera include, but are not limited to,armyworms, cutworms, loopers and heliothines in the family NoctuidaeSpodoptera frugiperda JE Smith (fall armyworm); S. exigua Hübner (beetarmyworm); S. litura Fabricius (tobacco cutworm, cluster caterpillar);Mamestra configurata Walker (bertha armyworm); M. brassicae Linnaeus(cabbage moth); Agrotis ipsilon Hufnagel (black cutworm); A. orthogoniaMorrison (western cutworm); A. subterranea Fabricius (granulatecutworm); Alabama argillacea Hübner (cotton leaf worm); Trichoplusia niHübner (cabbage looper); Pseudoplusia includens Walker (soybean looper);Anticarsia gemmatalis Hübner (velvetbean caterpillar); Hypena scabraFabricius (green cloverworm); Heliothis virescens Fabricius (tobaccobudworm); Pseudaletia unipuncta Haworth (armyworm); Athetis mindaraBarnes and Mcdunnough (rough skinned cutworm); Euxoa messoria Harris(darksided cutworm); Earias insulana Boisduval (spiny bollworm); E.vittella Fabricius (spotted bollworm); Helicoverpa armigera Hübner(American bollworm); H. zea Boddie (corn earworm or cotton bollworm);Melanchra picta Harris (zebra caterpillar); Egira (Xylomyges) curialisGrote (citrus cutworm); borers, casebearers, webworms, coneworms, andskeletonizers from the family Pyralidae Ostrinia nubilalis Hübner(European corn borer); Amyelois transitella Walker (naval orangeworm);Anagasta kuehniella Zeller (Mediterranean flour moth); Cadra cautellaWalker (almond moth); Chilo suppressalis Walker (rice stem borer); C.partellus, (sorghum borer); Corcyra cephalonica Stainton (rice moth);Crambus caliginosellus Clemens (corn root webworm); C. teterrellusZincken (bluegrass webworm); Cnaphalocrocis medinalis Guenée (rice leafroller); Desmia funeralis Hübner (grape leaffolder); Diaphania hyalinataLinnaeus (melon worm); D. nitidalis Stoll (pickleworm); Diatraeagrandiosella Dyar (southwestern corn borer), D. saccharalis Fabricius(surgarcane borer); Eoreuma loftini Dyar (Mexican rice borer); Ephestiaelutella Hübner (tobacco (cacao) moth); Galleria mellonella Linnaeus(greater wax moth); Herpetogramma licarsisalis Walker (sod webworm);Homoeosoma electellum Hulst (sunflower moth); Elasmopalpus lignosellusZeller (lesser cornstalk borer); Achroia grisella Fabricius (lesser waxmoth); Loxostege sticticalis Linnaeus (beet webworm); Orthaga thyrisalisWalker (tea tree web moth); Maruca testulalis Geyer (bean pod borer);Plodia interpunctella Hübner (Indian meal moth); Scirpophaga incertulasWalker (yellow stem borer); Udea rubigalis Guenée (celery leaf tier);and leaf rollers, budworms, seed worms and fruit worms in the familyTortricidae Acleris gloverana Walsingham (Western blackheaded budworm);A. variana Fernald (Eastern blackheaded budworm); Archips argyrospilaWalker (fruit tree leaf roller); A. rosana Linnaeus (European leafroller); and other Archips species, Adoxophyes orana Fischer vonRösslerstamm (summer fruit tortrix moth); Cochylis hospes Walsing ham(banded sunflower moth); Cydia latiferreana Walsing ham (filbertworm);C. pomonella Linnaeus (coding moth); Platynota flavedana Clemens(variegated leafroller); P. stultana Walsingham (omnivorous leafroller);Lobesia botrana Denis & Schiffermüller (European grape vine moth);Spilonota ocellana Denis & Schiffermüller (eyespotted bud moth);Endopiza viteana Clemens (grape berry moth); Eupoecilia ambiguellaHübner (vine moth); Bonagota salubricola Meyrick (Brazilian appleleafroller); Grapholita molesta Busck (oriental fruit moth); Suleimahelianthana Riley (sunflower bud moth); Argyrotaenia spp.; andChoristoneura spp.

Selected other agronomic pests in the order Lepidoptera include, but arenot limited to, Alsophila pometaria Harris (fall cankerworm); Anarsialineatella Zeller (peach twig borer); Anisota senatoria J. E. Smith(orange striped oakworm); Antheraea pernyi Guérin-Meneville (Chinese OakTussah Moth); Bombyx mori Linnaeus (Silkworm); Bucculatrix thurberiellaBusck (cotton leaf perforator); Colias eurytheme Boisduval (alfalfacaterpillar); Datana integerrima Grote & Robinson (walnut caterpillar);Dendrolimus sibiricus Tschetwerikov (Siberian silk moth), Ennomossubsignaria Hübner (elm spanworm); Erannis tiliaria Harris (lindenlooper); Euproctis chrysorrhoea Linnaeus (browntail moth); Harrisinaamericana Guérin-Meneville (grapeleaf skeletonizer); Hemileuca oliviaeCockrell (range caterpillar); Hyphantria cunea Drury (fall webworm);Keiferia lycopersicella Walsingham (tomato pinworm); Lambdinafiscellaria fiscellaria Hulst (Eastern hemlock looper); L. fiscellarialugubrosa Hulst (Western hemlock looper); Leucoma salicis Linnaeus(satin moth); Lymantria dispar Linnaeus (gypsy moth); Manducaquinquemaculata Haworth (five spotted hawk moth, tomato hornworm); M.sexta Haworth (tomato hornworm, tobacco hornworm); Operophtera brumataLinnaeus (winter moth); Paleacrita vernata Peck (spring cankerworm);Papilio cresphontes Cramer (giant swallowtail orange dog); Phryganidiacalifornica Packard (California oakworm); Phyllocnistis citrellaStainton (citrus leafminer); Phyllonorycter blancardella Fabricius(spotted tentiform leafminer); Pieris brassicae Linnaeus (large whitebutterfly); P. rapae Linnaeus (small white butterfly); P. napi Linnaeus(green veined white butterfly); Platyptilia carduidactyla Riley(artichoke plume moth); Plutella xylostella Linnaeus (diamondback moth);Pectinophora gossypiella Saunders (pink bollworm); Pontia protodiceBoisduval and Leconte (Southern cabbageworm); Sabulodes aegrotata Guenée(omnivorous looper); Schizura concinna J. E. Smith (red humpedcaterpillar); Sitotroga cerealella Olivier (Angoumois grain moth);Thaumetopoea pityocampa Schiffermuller (pine processionary caterpillar);Tineola bisselliella Hummel (webbing clothesmoth); Tuta absoluta Meyrick(tomato leafminer); Yponomeuta padella Linnaeus (ermine moth); Heliothissubflexa Guenée; Malacosoma spp. and Orgyia spp.

Of interest are larvae and adults of the order Coleoptera includingweevils from the families Anthribidae, Bruchidae and Curculionidae(including, but not limited to: Anthonomus grandis Boheman (bollweevil); Lissorhoptrus oryzophilus Kuschel (rice water weevil);Sitophilus granarius Linnaeus (granary weevil); S. oryzae Linnaeus (riceweevil); Hypera punctata Fabricius (clover leaf weevil);Cylindrocopturus adspersus LeConte (sunflower stem weevil); Smicronyxfulvus LeConte (red sunflower seed weevil); S. sordidus LeConte (graysunflower seed weevil); Sphenophorus maidis Chittenden (maize billbug));flea beetles, cucumber beetles, rootworms, leaf beetles, potato beetlesand leafminers in the family Chrysomelidae (including, but not limitedto: Leptinotarsa decemlineata Say (Colorado potato beetle); Diabroticavirgifera virgifera LeConte (western corn rootworm); D. barberi Smithand Lawrence (northern corn rootworm); D. undecimpunctata howardi Barber(southern corn rootworm); Chaetocnema pulicaria Melsheimer (corn fleabeetle); Phyllotreta cruciferaeGoeze (Crucifer flea beetle); Phyllotretastriolata (stripped flea beetle); Colaspis brunnea Fabricius (grapecolaspis); Oulema melanopus Linnaeus (cereal leaf beetle); Zygogrammaexclamationis Fabricius (sunflower beetle)); beetles from the familyCoccinellidae (including, but not limited to: Epilachna varivestisMulsant (Mexican bean beetle)); chafers and other beetles from thefamily Scarabaeidae (including, but not limited to: Popillia japonicaNewman (Japanese beetle); Cyclocephala borealis Arrow (northern maskedchafer, white grub); C. immaculata Olivier (southern masked chafer,white grub); Rhizotrogus majalis Razoumowsky (European chafer);Phyllophaga crinita Burmeister (white grub); Ligyrus gibbosus De Geer(carrot beetle)); carpet beetles from the family Dermestidae; wirewormsfrom the family Elateridae, Eleodes spp., Melanotus spp.; Conoderusspp.; Limonius spp.; Agriotes spp.; Ctenicera spp.; Aeolus spp.; barkbeetles from the family Scolytidae and beetles from the familyTenebrionidae.

Adults and immatures of the order Diptera are of interest, includingleafminers Agromyza parvicornis Loew (corn blotch leafminer); midges(including, but not limited to: Contarinia sorghicola Coquillett(sorghum midge); Mayetiola destructor Say (Hessian fly); Sitodiplosismosellana Géhin (wheat midge); Neolasioptera murtfeldtiana Felt,(sunflower seed midge)); fruit flies (Tephritidae), Oscinella fritLinnaeus (fruit flies); maggots (including, but not limited to: Deliaplatura Meigen (seedcorn maggot); D. coarctata Fallen (wheat bulb fly)and other Delia spp., Meromyza americana Fitch (wheat stem maggot);Musca domestica Linnaeus (house flies); Fannia canicularis Linnaeus, F.femoralis Stein (lesser house flies); Stomoxys calcitrans Linnaeus(stable flies)); face flies, horn flies, blow flies, Chrysomya spp.;Phormia spp. and other muscoid fly pests, horse flies Tabanus spp.; botflies Gastrophilus spp.; Oestrus spp.; cattle grubs Hypoderma spp.; deerflies Chrysops spp.; Melophagus ovinus Linnaeus (keds) and otherBrachycera, mosquitoes Aedes spp.; Anopheles spp.; Culex spp.; blackflies Prosimulium spp.; Simulium spp.; biting midges, sand flies,sciarids, and other Nematocera.

Included as insects of interest are adults and nymphs of the ordersHemiptera and Homoptera such as, but not limited to, adelgids from thefamily Adelgidae, plant bugs from the family Miridae, cicadas from thefamily Cicadidae, leafhoppers, Empoasca spp.; from the family

Cicadellidae, planthoppers from the families Cixiidae, Flatidae,Fulgoroidea, lssidae and Delphacidae, treehoppers from the familyMembracidae, psyllids from the family Psyllidae, whiteflies from thefamily Aleyrodidae, aphids from the family Aphididae, phylloxera fromthe family Phylloxeridae, mealybugs from the family Pseudococcidae,scales from the families Asterolecanidae, Coccidae, Dactylopiidae,Diaspididae, Eriococcidae Ortheziidae,

Phoenicococcidae and Margarodidae, lace bugs from the family Tingidae,stink bugs from the family Pentatomidae, cinch bugs, Blissus spp.; andother seed bugs from the family Lygaeidae, spittlebugs from the familyCercopidae squash bugs from the family Coreidae and red bugs and cottonstainers from the family Pyrrhocoridae.

Agronomically important members from the order Homoptera furtherinclude, but are not limited to: Acyrthisiphon pisum Harris (pea aphid);Aphis craccivora Koch (cowpea aphid); A. fabae Scopoli (black beanaphid); A. gossypii Glover (cotton aphid, melon aphid); A. maidiradicisForbes (corn root aphid); A. pomi De Geer (apple aphid); A. spiraecolaPatch (spirea aphid); Aulacorthum solani Kaltenbach (foxglove aphid);Chaetosiphon fragaefolii Cockerell (strawberry aphid); Diuraphis noxiaKurdjumov/Mordvilko (Russian wheat aphid); Dysaphis plantagineaPaaserini (rosy apple aphid); Eriosoma lanigerum Hausmann (woolly appleaphid); Brevicoryne brassicae Linnaeus (cabbage aphid); Hyalopteruspruni Geoffroy (mealy plum aphid); Lipaphis erysimi Kaltenbach (turnipaphid); Metopolophium dirrhodum Walker (cereal aphid); Macrosiphumeuphorbiae Thomas (potato aphid); Myzus persicae Sulzer (peach-potatoaphid, green peach aphid); Nasonovia ribisnigri Mosley (lettuce aphid);Pemphigus spp. (root aphids and gall aphids); Rhopalosiphum maidis Fitch(corn leaf aphid); R. padi Linnaeus (bird cherry-oat aphid); Schizaphisgraminum Rondani (greenbug); Sipha flava Forbes (yellow sugarcaneaphid); Sitobion avenae Fabricius (English grain aphid); Therioaphismaculata Buckton (spotted alfalfa aphid); Toxoptera aurantii Boyer deFonscolombe (black citrus aphid) and T. citricida Kirkaldy (brown citrusaphid); Adelges spp. (adelgids); Phylloxera devastatrix Pergande (pecanphylloxera); Bemisia tabaci Gennadius (tobacco whitefly, sweetpotatowhitefly); B. argentifolii Bellows & Perring (silverleaf whitefly);Dialeurodes citri Ashmead (citrus whitefly); Trialeurodes abutiloneus(bandedwinged whitefly) and T. vaporariorum Westwood (greenhousewhitefly); Empoasca fabae Harris (potato leafhopper); Laodelphaxstriatellus Fallen (smaller brown planthopper); Macrolestesquadrilineatus Forbes (aster leafhopper); Nephotettix cinticeps Uhler(green leafhopper); N. nigropictus Stål (rice leafhopper); Nilaparvatalugens Stål (brown planthopper); Peregrinus maidis Ashmead (cornplanthopper); Sogatella furcifera Horvath (white-backed planthopper);Sogatodes orizicola Muir (rice delphacid); Typhlocyba pomaria McAtee(white apple leafhopper); Erythroneoura spp. (grape leafhoppers);Magicicada septendecim Linnaeus (periodical cicada); Icerya purchasiMaskell (cottony cushion scale); Quadraspidiotus perniciosus Comstock(San Jose scale); Planococcus citri Risso (citrus mealybug);Pseudococcus spp. (other mealybug complex); Cacopsylla pyricola Foerster(pear psylla); Trioza diospyri Ashmead (persimmon psylla).

Agronomically important species of interest from the order Hemipterainclude, but are not limited to: Acrosternum hilare Say (green stinkbug); Anasa tristis De Geer (squash bug); Blissus leucopterusleucopterus Say (chinch bug); Corythuca gossypii Fabricius (cotton lacebug); Cyrtopeltis modesta Distant (tomato bug); Dysdercus suturellusHerrich-Schaffer (cotton stainer); Euschistus servus Say (brown stinkbug); E. variolarius Palisot de Beauvois (one-spotted stink bug);Graptostethus spp. (complex of seed bugs); Leptoglossus corculus Say(leaf-footed pine seed bug); Lygus lineolaris Palisot de Beauvois(tarnished plant bug); L. Hesperus Knight (Western tarnished plant bug);L. pratensis Linnaeus (common meadow bug); L. rugulipennis Poppius(European tarnished plant bug); Lygocoris pabulinus Linnaeus (commongreen capsid); Nezara viridula Linnaeus (southern green stink bug);Oebalus pugnax Fabricius (rice stink bug); Oncopeltus fasciatus Dallas(large milkweed bug); Pseudatomoscelis seriatus Reuter (cottonfleahopper).

Furthermore, embodiments may be effective against Hemiptera such,Calocoris norvegicus Gmelin (strawberry bug); Orthops campestrisLinnaeus; Plesiocoris rugicollis Fallen (apple capsid); Cyrtopeltismodestus Distant (tomato bug); Cyrtopeltis notatus Distant (suckfly);Spanagonicus albofasciatus Reuter (whitemarked fleahopper); Diaphnocorischlorionis Say (honeylocust plant bug); Labopidicola allii Knight (onionplant bug); Pseudatomoscelis seriatus Reuter (cotton fleahopper);Adelphocoris rapidus Say (rapid plant bug); Poecilocapsus lineatusFabricius (four-lined plant bug); Nysius ericae Schilling (false chinchbug); Nysius raphanus Howard (false chinch bug); Nezara viridulaLinnaeus (Southern green stink bug); Eurygaster spp.; Coreidae spp.;Pyrrhocoridae spp.; Tinidae spp.; Blostomatidae spp.; Reduviidae spp.and Cimicidae spp.

Also included are adults and larvae of the order Acari (mites) such asAceria tosichella Keifer (wheat curl mite); Petrobia latens Muller(brown wheat mite); spider mites and red mites in the familyTetranychidae, Panonychus ulmi Koch (European red mite); Tetranychusurticae Koch (two spotted spider mite); (T. mcdanieli McGregor (McDanielmite); T. cinnabarinus Boisduval (carmine spider mite); T. turkestaniUgarov & Nikolski (strawberry spider mite); flat mites in the familyTenuipalpidae, Brevipalpus lewisi McGregor (citrus flat mite); rust andbud mites in the family Eriophyidae and other foliar feeding mites andmites important in human and animal health, i.e., dust mites in thefamily Epidermoptidae, follicle mites in the family Demodicidae, grainmites in the family Glycyphagidae, ticks in the order Ixodidae. Ixodesscapularis Say (deer tick); I. holocyclus Neumann (Australian paralysistick); Dermacentor variabilis Say (American dog tick); Amblyommaamericanum Linnaeus (lone star tick) and scab and itch mites in thefamilies Psoroptidae, Pyemotidae and Sarcoptidae.

Insect pests of the order Thysanura are of interest, such as Lepismasaccharina Linnaeus (silverfish); Thermobia domestica Packard(firebrat).

Additional arthropod pests covered include: spiders in the order Araneaesuch as Loxosceles reclusa Gertsch and Mulaik (brown recluse spider) andthe Latrodectus mactans Fabricius (black widow spider) and centipedes inthe order Scutigeromorpha such as Scutigera coleoptrata Linnaeus (housecentipede).

Insect pest of interest include the superfamily of stink bugs and otherrelated insects including but not limited to species belonging to thefamily Pentatomidae (Nezara viridula, Halyomorpha halys, Piezodorusguildini, Euschistus servus, Acrosternum hilare, Euschistus heros,Euschistus tristigmus, Acrosternum hilare, Dichelops furcatus, Dichelopsmelacanthus, and Bagrada hilaris (Bagrada Bug)), the family Plataspidae(Megacopta cribraria—Bean plataspid) and the family Cydnidae(Scaptocoris castanea—Root stink bug) and Lepidoptera species includingbut not limited to: diamond-back moth, e.g., Helicoverpa zea Boddie;soybean looper, e.g., Pseudoplusia includens Walker and velvet beancaterpillar e.g., Anticarsia gemmatalis Hübner.

Methods for measuring pesticidal activity are well known in the art.See, for example, Czapla and Lang, (1990) J. Econ. EntomoL 83:2480-2485;Andrews, et al., (1988) Biochem. J. 252:199-206; Marrone, et al., (1985)J. of Economic Entomology 78:290-293 and U.S. Pat. No. 5,743,477, all ofwhich are herein incorporated by reference in their entirety. Generally,the protein is mixed and used in feeding assays. See, for exampleMarrone, et al., (1985) J. of Economic Entomology 78:290-293. Suchassays can include contacting plants with one or more pests anddetermining the plant's ability to survive and/or cause the death of thepests.

Nematodes include parasitic nematodes such as root-knot, cyst and lesionnematodes, including Heterodera spp., Meloidogyne spp. and Globoderaspp.; particularly members of the cyst nematodes, including, but notlimited to, Heterodera glycines (soybean cyst nematode); Heteroderaschachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode)and Globodera rostochiensis and Globodera pailida (potato cystnematodes). Lesion nematodes include Pratylenchus spp.

Seed Treatment

To protect and to enhance yield production and trait technologies, seedtreatment options can provide additional crop plan flexibility and costeffective control against insects, weeds and diseases. Seed material canbe treated, typically surface treated, with a composition comprisingcombinations of chemical or biological herbicides, herbicide safeners,insecticides, fungicides, germination inhibitors and enhancers,nutrients, plant growth regulators and activators, bactericides,nematocides, avicides and/or molluscicides. These compounds aretypically formulated together with further carriers, surfactants orapplication-promoting adjuvants customarily employed in the art offormulation. The coatings may be applied by impregnating propagationmaterial with a liquid formulation or by coating with a combined wet ordry formulation. Examples of the various types of compounds that may beused as seed treatments are provided in The Pesticide Manual: A WorldCompendium, C.D.S. Tomlin Ed., Published by the British Crop ProductionCouncil, which is hereby incorporated by reference.

Some seed treatments that may be used on crop seed include, but are notlimited to, one or more of abscisic acid, acibenzolar-S-methyl,avermectin, amitrol, azaconazole, azospirillum, azadirachtin,azoxystrobin, Bacillus spp. (including one or more of cereus, firmus,megaterium, pumilis, sphaericus, subtilis and/or thuringiensis species),bradyrhizobium spp. (including one or more of betae, canariense,elkanii, iriomotense, japonicum, liaonigense, pachyrhizi and/oryuanmingense), captan, carboxin, chitosan, clothianidin, copper,cyazypyr, difenoconazole, etidiazole, fipronil, fludioxonil,fluoxastrobin, fluquinconazole, flurazole, fluxofenim, harpin protein,imazalil, imidacloprid, ipconazole, isoflavenoids,lipo-chitooligosaccharide, mancozeb, manganese, maneb, mefenoxam,metalaxyl, metconazole, myclobutanil, PCNB, penflufen, penicillium,penthiopyrad, permethrine, picoxystrobin, prothioconazole,pyraclostrobin, rynaxypyr, S-metolachlor, saponin, sedaxane, TCMTB,tebuconazole, thiabendazole, thiamethoxam, thiocarb, thiram,tolclofos-methyl, triadimenol, trichoderma, trifloxystrobin,triticonazole and/or zinc. PCNB seed coat refers to EPA RegistrationNumber 00293500419, containing quintozen and terrazole. TCMTB refers to2-(thiocyanomethylthio) benzothiazole.

Seed varieties and seeds with specific transgenic traits may be testedto determine which seed treatment options and application rates maycomplement such varieties and transgenic traits to enhance yield. Forexample, a variety with good yield potential but head smutsusceptibility may benefit from the use of a seed treatment thatprovides protection against head smut, a variety with good yieldpotential but cyst nematode susceptibility may benefit from the use of aseed treatment that provides protection against cyst nematode, and soon. Likewise, a variety encompassing a transgenic trait conferringinsect resistance may benefit from the second mode of action conferredby the seed treatment, a variety encompassing a transgenic traitconferring herbicide resistance may benefit from a seed treatment with asafener that enhances the plants resistance to that herbicide, etc.Further, the good root establishment and early emergence that resultsfrom the proper use of a seed treatment may result in more efficientnitrogen use, a better ability to withstand drought and an overallincrease in yield potential of a variety or varieties containing acertain trait when combined with a seed treatment.

Methods for Killing an Insect Pest and Controlling an Insect Population

In some embodiments, methods are provided for killing an insect pest,comprising contacting the insect pest, either simultaneously orsequentially, with an insecticidally-effective amount of a recombinantIPD113 polypeptide or IPD113 chimeric polypeptide of the disclosure. Insome embodiments, methods are provided for killing an insect pest,comprising contacting the insect pest with an insecticidally-effectiveamount of a recombinant pesticidal protein of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7,SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12,SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ IDNO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31,SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO:36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ IDNO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO:55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ IDNO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69,SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO:74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ IDNO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88,SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO:93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ IDNO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102,SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ IDNO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111,SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ IDNO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120,SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ IDNO: 125, SEQ ID NO: 126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255,SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ IDNO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264,SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ IDNO: 269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273,SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ IDNO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311,SEQ ID NO: 312, SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ IDNO: 316, SEQ ID NO: 317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320,SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ IDNO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427,SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ IDNO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436,SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ IDNO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450,SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ IDNO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469,SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ IDNO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480,SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ IDNO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489,SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ IDNO: 495 or a variant thereof.

In some embodiments, methods are provided for controlling an insect pestpopulation, comprising contacting the insect pest population, eithersimultaneously or sequentially, with an insecticidally-effective amountof a recombinant IPD113 polypeptide or IPD113 chimeric polypeptide ofthe disclosure. In some embodiments, methods are provided forcontrolling an insect pest population, comprising contacting the insectpest population with an insecticidally-effective amount of a recombinantIPD113 polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 or avariant thereof. As used herein, “controlling a pest population” or“controls a pest” refers to any effect on a pest that results inlimiting the damage that the pest causes. Controlling a pest includes,but is not limited to, killing the pest, inhibiting development of thepest, altering fertility or growth of the pest in such a manner that thepest provides less damage to the plant, decreasing the number ofoffspring produced, producing less fit pests, producing pests moresusceptible to predator attack or deterring the pests from eating theplant.

In some embodiments, methods are provided for controlling an insect pestpopulation resistant to a pesticidal protein, comprising contacting theinsect pest population, either simultaneously or sequentially, with aninsecticidally-effective amount of a recombinant IPD113 polypeptide orchimeric IPD113 polypeptide of the disclosure. In some embodiments,methods are provided for controlling an insect pest population resistantto a pesticidal protein, comprising contacting the insect pestpopulation with an insecticidally-effective amount of a recombinantIPD113 polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 or avariant thereof.

In some embodiments, methods are provided for protecting a plant from aninsect pest, comprising expressing in the plant or cell thereof at leastone recombinant polynucleotide encoding an IPD113 polypeptide orchimeric IPD113 polypeptide. In some embodiments, methods are providedfor protecting a plant from an insect pest, comprising expressing in theplant or cell thereof a recombinant polynucleotide encoding IPD113polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO:33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ IDNO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47,SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO:52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ IDNO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66,SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO:71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ IDNO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85,SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO:90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ IDNO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO:104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO:113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO:122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO:257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261, SEQID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO:266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQ ID NO:275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQ ID NO:313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQ ID NO:419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO:429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO:439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO:452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO:472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO:482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO:491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 or variantsthereof.

Insect Resistance Management (IRM) Strategies

Expression of B. thuringiensis δ-endotoxins in transgenic corn plantshas proven to be an effective means of controlling agriculturallyimportant insect pests (Perlak, et al., 1990; 1993). However, insectshave evolved that are resistant to B. thuringiensis δ-endotoxinsexpressed in transgenic plants. Such resistance, should it becomewidespread, would clearly limit the commercial value of germplasmcontaining genes encoding such B. thuringiensis δ-endotoxins.

One way to increasing the effectiveness of the transgenic insecticidesagainst target pests and contemporaneously reducing the development ofinsecticide-resistant pests is to use provide non-transgenic (i.e.,non-insecticidal protein) refuges (a section of non-insecticidalcrops/corn) for use with transgenic crops producing a singleinsecticidal protein active against target pests. The United StatesEnvironmental Protection Agency(epa.gov/oppbppdl/biopesticides/pips/bt_corn_refuge_2006.htm, which canbe accessed using the www prefix) publishes the requirements for usewith transgenic crops producing a single Bt protein active againsttarget pests. In addition, the National Corn Growers Association, ontheir website:(ncga.com/insect-resistance-management-fact-sheet-bt-corn, which can beaccessed using the www prefix) also provides similar guidance regardingrefuge requirements. Due to losses to insects within the refuge area,larger refuges may reduce overall yield.

Another way of increasing the effectiveness of the transgenicinsecticides against target pests and contemporaneously reducing thedevelopment of insecticide-resistant pests would be to have a repositoryof insecticidal genes that are effective against groups of insect pestsand which manifest their effects through different modes of action.

Expression in a plant of two or more insecticidal compositions toxic tothe same insect species, each insecticide being expressed at efficaciouslevels would be another way to achieve control of the development ofresistance. This is based on the principle that evolution of resistanceagainst two separate modes of action is far more unlikely than only one.Roush, for example, outlines two-toxin strategies, also called“pyramiding” or “stacking,” for management of insecticidal transgeniccrops. (The Royal Society. Phil. Trans. R. Soc. Lond. B. (1998)353:1777-1786). Stacking or pyramiding of two different proteins eacheffective against the target pests and with little or nocross-resistance can allow for use of a smaller refuge. The USEnvironmental Protection Agency requires significantly less (generally5%) structured refuge of non-Bt corn be planted than for single traitproducts (generally 20%). There are various ways of providing the IRMeffects of a refuge, including various geometric planting patterns inthe fields and in-bag seed mixtures, as discussed further by Roush.

In some embodiments, the IPD113 polypeptides of the disclosure areuseful as an insect resistance management strategy in combination (i.e.,pyramided) with other pesticidal proteins include but are not limited toBt toxins, Xenorhabdus sp. or Photorhabdus sp. insecticidal proteins,other insecticidally active proteins, and the like.

Provided are methods of controlling Lepidoptera and/or Coleoptera insectinfestation(s) in a transgenic plant that promote insect resistancemanagement, comprising expressing in the plant at least two differentinsecticidal proteins having different modes of action.

In some embodiments, the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management comprises the presentation of at least one of theIPD113 polypeptide insecticidal proteins to insects in the orderLepidoptera and/or Coleoptera.

In some embodiments, the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management comprises the presentation of at least one of theIPD113 polypeptides of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319 or SEQ ID NO: 320 or variantsthereof, insecticidal to insects in the order Lepidoptera and/orColeoptera.

In some embodiments, the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management comprise expressing in the transgenic plant anIPD113 polypeptide and a Cry protein or other insecticidal protein toinsects in the order Lepidoptera and/or Coleoptera having differentmodes of action.

In some embodiments, the methods, of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management, comprise expression in the transgenic plant anIPD113 polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 or variantsthereof and a Cry protein or other insecticidal protein to insects inthe order Lepidoptera and/or Coleoptera, where the IPD113 polypeptideand Cry protein have different modes of action.

Also provided are methods of reducing likelihood of emergence ofLepidoptera and/or Coleoptera insect resistance to transgenic plantsexpressing in the plants insecticidal proteins to control the insectspecies, comprising expression of an IPD113 polypeptide insecticidal tothe insect species in combination with a second insecticidal protein tothe insect species having different modes of action.

Also provided are means for effective Lepidoptera and/or Coleopterainsect resistance management of transgenic plants, comprisingco-expressing at high levels in the plants two or more insecticidalproteins toxic to Lepidoptera and/or Coleoptera insects but eachexhibiting a different mode of effectuating its killing activity,wherein the two or more insecticidal proteins comprise an IPD113polypeptide and a Cry protein. Also provided are means for effectiveLepidoptera and/or Coleoptera insect resistance management of transgenicplants, comprising co-expressing at high levels in the plants two ormore insecticidal proteins toxic to Lepidoptera and/or Coleopterainsects but each exhibiting a different mode of effectuating its killingactivity, wherein the two or more insecticidal proteins comprise anIPD113 polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 or variantsthereof and a Cry protein or other insecticidally active protein.

In addition, methods are provided for obtaining regulatory approval forplanting or commercialization of plants expressing proteins insecticidalto insects in the order Lepidoptera and/or Coleoptera, comprising thestep of referring to, submitting or relying on insect assay binding datashowing that the IPD113 polypeptide does not compete with binding sitesfor Cry proteins in such insects. In addition, methods are provided forobtaining regulatory approval for planting or commercialization ofplants expressing proteins insecticidal to insects in the orderLepidoptera and/or Coleoptera, comprising the step of referring to,submitting or relying on insect assay binding data showing that theIPD113 polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ IDNO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO:42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ IDNO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO:61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ IDNO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75,SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO:117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:126, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO:261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO:270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274, SEQID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO:279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 311, SEQ ID NO: 312, SEQID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO:317, SEQ ID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 416, SEQID NO: 419, SEQ ID NO: 420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO:424, SEQ ID NO: 425, SEQ ID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQID NO: 429, SEQ ID NO: 430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO:433, SEQ ID NO: 434, SEQ ID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQID NO: 439, SEQ ID NO: 440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO:445, SEQ ID NO: 447, SEQ ID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQID NO: 452, SEQ ID NO: 453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO:461, SEQ ID NO: 463, SEQ ID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQID NO: 472, SEQ ID NO: 473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO:477, SEQ ID NO: 478, SEQ ID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQID NO: 482, SEQ ID NO: 483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO:486, SEQ ID NO: 487, SEQ ID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQID NO: 491, SEQ ID NO: 492, SEQ ID NO: 494 or SEQ ID NO: 495 or variantthereof does not compete with binding sites for Cry proteins in suchinsects.

Methods for Increasing Plant Yield

Methods for increasing plant yield are provided. The methods compriseproviding a plant or plant cell expressing a polynucleotide encoding thepesticidal polypeptide sequence disclosed herein and growing the plantor a seed thereof in a field infested with a pest against which thepolypeptide has pesticidal activity. In some embodiments, thepolypeptide has pesticidal activity against a Lepidopteran, Coleopteran,Dipteran, Hemipteran or nematode pest, and the field is infested with aLepidopteran, Hemipteran, Coleopteran, Dipteran or nematode pest.

As defined herein, the “yield” of the plant refers to the quality and/orquantity of biomass produced by the plant. “Biomass” as used hereinrefers to any measured plant product. An increase in biomass productionis any improvement in the yield of the measured plant product.Increasing plant yield has several commercial applications. For example,increasing plant leaf biomass may increase the yield of leafy vegetablesfor human or animal consumption. Additionally, increasing leaf biomasscan be used to increase production of plant-derived pharmaceutical orindustrial products. An increase in yield can comprise any statisticallysignificant increase including, but not limited to, at least a 1%increase, at least a 3% increase, at least a 5% increase, at least a 10%increase, at least a 20% increase, at least a 30%, at least a 50%, atleast a 70%, at least a 100% or a greater increase in yield compared toa plant not expressing the pesticidal sequence.

In specific methods, plant yield is increased as a result of improvedpest resistance of a plant expressing an IPD113 polypeptide disclosedherein. Expression of the IPD113 polypeptide results in a reducedability of a pest to infest or feed on the plant, thus improving plantyield.

Methods of Processing

Further provided are methods of processing a plant, plant part or seedto obtain a food or feed product from a plant, plant part or seedcomprising an IPD113 polynucleotide. The plants, plant parts or seedsprovided herein, can be processed to yield oil, protein products and/orby-products that are derivatives obtained by processing that havecommercial value. Non-limiting examples include transgenic seedscomprising a nucleic acid molecule encoding an IPD113 polypeptide whichcan be processed to yield soy oil, soy products and/or soy by-products.

“Processing” refers to any physical and chemical methods used to obtainany soy product and includes, but is not limited to, heat conditioning,flaking and grinding, extrusion, solvent extraction or aqueous soakingand extraction of whole or partial seeds

The following examples are offered by way of illustration and not by wayof limitation.

EXAMPLES Example 1—Isolation and Identification of an InsecticidalProtein Active Against Lepidoptera species from the Fern, Pteris cretica

Insecticidal activity against soybean looper ((SBL) (Pseudoplusiaincludens)) and corn earworm ((CEW) (Helicoverpa zea)) was observed froma clarified and desalted extraction from Pteris cretica cv albolineata(PS930) and Pteris umbrosa (PS995) plant tissue. This insecticidalactivity exhibited heat and protease sensitivity indicatingproteinaceous nature.

Pteris cretica cv albolineata (PS930) and Pteris umbrosa (PS995) hadsimilar activity profiles from crude sample and anion exchange separatedfractions and similar SDS-PAGE protein profiles at the crude level. Dueto the limited amount of material the samples were combined into onesample for the purification steps.

The PS930 and PS995 combined plant material was removed from storage at−80° C. and ground to a fine powder at liquid Nitrogen temperatures witha Geno/Grinder® 2010 Ball Mill (SPEX Sample Prep®, Metuchen, N.J.). Theprotein was extracted from the plant tissue by adding extraction buffer((50 mM Tris, pH 8.0, 150 mM Potassium Chloride, 2.5 mM EDTA, 1.5%Polyvinylpolypyrrolidone and Complete EDTA Free protease inhibitortablets (Roche Diagnostics, Germany)) at a ratio of four mL per everyone gram of fresh weight of tissue. The sample was kept in suspension bylight agitation on a platform rocker at 4° C. for 15 minutes. Thehomogenate was clarified by centrifugation at 6000×g for 15 minutesfollowed by filtration through a Whatman 0.45 μm filter (GE Healthcare,Piscataway, N.J.). PS930 and PS995 were desalted into 50 mM Tris, pH 8.0using 10 mL Zeba™ Spin desalting columns (Thermo Scientific, Ill.)before loading onto a 5 mL HiTrap™Q-FF column (GE Healthcare,Piscataway, N.J.) that was equilibrated in the same buffer. A linear 30column volume gradient from 0.0 M to 0.7 M NaCl in 50 mM Tris, pH 8.0was used to elute bound protein. The eluted fractions and flow-throughwere assayed against SBL as described in Example 6. Activity against SBLwas detected in fractions eluting at approximately 13.0-31.0 mS/cm². Thefractions were pooled and concentrated 10× on a 3 kDa MWCO filter (PallLife Sciences, Port Washington, N.Y.) and loaded onto a HiPrep™ 16/60Superdex 300 size exclusion column (GE Healthcare, Piscataway, N.J.). Anisocratic gradient of 50 mM Tris, pH 8.0 was applied and the eluted 1 mLfractions were assayed against SBL and CEW. The active fractions werecombined and injected onto a 5 mL Mono Q® 5/50 column (GE Healthcare,Piscataway, N.J.) equilibrated in 50 mM Tris, pH 8.0. A 65-column volumelinear gradient from 0% to 60% Elution Buffer (50 mM Tris pH 8.0, 1.0 MNaCl) was performed to generate 0.5 mL fractions of eluted protein. Theeluted proteins were bioassayed as previously described and SBL and CEWactivity was detected in fractions eluting at ˜7.5-12.7 mS/cm²conductivity. The active fractions were concentrated individually 10×and run on a LDS-PAGE and individual bands were excised for in geldigest.

Proteins for MS identification were obtained after running the sample onan LDS-PAGE gel stained with Coomassie™ Brilliant Blue G-250 stain. Thebands of interest were excised from the gel, de-stained, reduced withdithiothreitol and then alkylated with iodoacetamide. Followingovernight digestion with trypsin, liquid chromatography-tandem massspectrometry (LC-MSMS) analysis for tryptically-digested peptides wasconducted using electrospray ion source on a QToF Premiere™ massspectrometer (Waters®, Milford, Mass.) coupled with a NanoAcquity™nano-LC system (Waters®, Milford, Mass.) with a gradient from 2%acetonitrile, 0.1% formic acid to 60% acetonitrile, 0.1% formic acid.

Protein identification was performed by database searches using Mascot®(Matrix Science, 10 Perrins Lane, London NW3 1QY UK). The searches wereconducted against an in-house transcriptome database containingtranscripts from the Pteris cretica cv albolineata (PS930), Pterisumbrosa (PS995) and other source plants and the public protein databaseSwiss-Prot using the Mascot search engine (Matrix Science). Proteinidentification was also performed by taking the resulting LCMS datawhich was analyzed using Protein Lynx Global Server™ (Waters®, Milford,Mass.) to generate DeNovo sequence data. The amino acid sequences wereBLAST™ (Basic Local Alignment Search Tool; Altschul, et al., (1993) J.Mol. Biol. 215:403-410; see also ncbi.nlm.nih.gov/BLAST/, which can beaccessed using the www prefix) searched against public andDUPONT-PIONEER internal databases that included plant protein sequences.Amino acid sequences were aligned with proteins in a proprietaryDUPONT-PIONEER plant protein database. Amino acid sequence from a bandof interest aligned with predicted protein from PS930.

Example 2—Transcriptomic Sequencing of Pteris cretica cv albolineata andCloning of IPD113Aa

A transcriptome for Pteris albolineata syn Pteris cretica cvalbolineata, (ID #PS930) was prepared as follows. Total RNAs wereisolated from frozen tissues by use of the Qiagen® RNeasy® kit for totalRNA isolation. Sequencing libraries from the resulting total RNAs wereprepared using the TruSeq™ mRNA-Seq kit and protocol from Illumina®,Inc. (San Diego, Calif.). Briefly, mRNAs were isolated via attachment tooligo(dT) beads, fragmented to a mean size of 180 nt, reversetranscribed into cDNA by random hexamer prime, end repaired, 3′A-tailed, and ligated with Illumina® indexed TruSeq™ adapters. LigatedcDNA fragments were PCR amplified using Illumina® TruSeq™ primers andpurified PCR products were checked for quality and quantity on theAgilent Bioanalyzer® DNA 7500 chip. Post quality and quantityassessment, 100 ng of the transcript library was normalized by treatmentwith Duplex-Specific Nuclease (DSN) (Evrogen®, Moscow, Russia).Normalization was accomplished by addition of 200 mM HEPES buffer,followed by heat denaturation and five hour anneal at 68° C. Annealedlibrary was treated with 2 ul of DSN enzyme for 25 minutes, purified byQiagen® MinElute® columns according to manufacturer protocols, andamplified twelve cycles using Illumina® adapter specific primers. Finalproducts were purified with Ampure® XP beads (Beckman Genomics, Danvers,Mass.) and checked for quality and quantity on the Agilent Bioanalyzer®DNA 7500 chip.

Normalized transcript libraries were sequenced according to manufacturerprotocols on the Illumina® Genome Analyzer IIx. Each library washybridized to two flowcell lanes and amplified, blocked, linearized andprimer hybridized using the Illumina clonal cluster generation processon cBot®. Sequencing was completed on the Genome Analyzer IIx,generating sixty million 75 bp paired end reads per normalized library.

Peptide sequences identified for IPD113Aa (SEQ ID NO: 1) by LCMSsequencing (described in Example 1) were searched against proteinsequences predicted by open reading frames (ORFs) from the transcriptomeassemblies for PS930. The peptides matched a transcript corresponding toIPD113Aa (SEQ ID NO: 1). The coding sequence was used to design thefollowing primers:cgaaatctctcatctaagaggctggatcctaggATGGATTCCGATCTGATTGCTCAG (SEQ ID NO:332) and gttggccaatccagaagatggacaagtctagaTCATGATGAGGGATCTTCAGGTG (SEQ IDNO: 333) to clone the IPD113Aa polynucleotide sequence (SEQ ID NO: 127)into a transient expression vector for expression and activity analysis.

Example 3—Expression and Insect Bioassay of IPD113Aa on Transient LeafTissues

To confirm activity of the IPD113Aa polypeptide (SEQ ID NO: 1) thecorresponding gene (SEQ ID NO: 127) was cloned into a transientexpression system under control of the viral promoter dMMV (Dey, et.al., (1999) Plant Mol. Biol. 40:771-782). The Agrobacterium strainscontaining the IPD113Aa expression construct was infiltrated intoleaves. The agro-infiltration method of introducing an Agrobacteriumcell suspension to plant cells of intact tissues so that reproducibleinfection and subsequent plant derived transgene expression may bemeasured or studied is well known in the art (Kapila, et. al., (1997)Plant Science 122:101-108). Briefly, the unifoliate leaves of bush bean(common bean, Phaseolus vulgaris) were agro-infiltrated with normalizedbacterial cell cultures of test and control strains. Leaf discs wereexcised from each plantlet and infested with 2 neonates of Soy BeanLooper (SBL) (Pseudoplusia includes), 2 neonates of Fall Armyworm (FAW)(Spodoptera frugiperda), 1 neonate of Corn Earworm (CEW) (Helicoverpazea), 3 neonates of Velvet Bean Caterpillar (VBC) (Anticarsiagemmatalis) or 3 neonates of European Corn Borer (ECB) (Ostrinianubialis). Leaf discs from a control were generated with Agrobacteriumnot containing an expression vector. Leaf discs from a non-infiltratedplant were used as a second control. The consumption of the leaf tissuewas scored three days after infestation (Table 1) and given scores of 0to 9 as indicated by Table 2.

TABLE 1 SBL FAW CEW ECB VBC Avg Std. Avg Std. Avg Std. Avg Std. Avg Std.Score DEV Score DEV Score DEV Score DEV Score DEV IPD113Aa 7.8 0.5 4.01.4 4.8 1.3 8.0 0.8 7.8 0.5 Empty Agro 1.3 0.5 1.0 0.0 1.3 0.5 1.3 0.51.0 0.0 Untreated 2.5 3.0 1.0 0.0 2.3 2.5 3.3 1.9 1.0 0.0

TABLE 1 SBL FAW CEW ECB VBC Avg Std. Avg Std. Avg Std. Avg Std. Avg Std.Score DEV Score DEV Score DEV Score DEV Score DEV IPD113Aa 7.8 0.5 4.01.4 4.8 1.3 8.0 0.8 7.8 0.5 Empty Agro 1.3 0.5 1.0 0.0 1.3 0.5 1.3 0.51.0 0.0 Untreated 2.5 3.0 1.0 0.0 2.3 2.5 3.3 1.9 1.0 0.0

Example 4—Identification of IPD113Aa Homologs

Gene identities may be determined by conducting BLAST™ (Basic LocalAlignment Search Tool; Altschul, et al., (1993) J. Mol. Biol.215:403-410; see also ncbi.nlm.nih.gov/BLAST/, which can be accessedusing the www prefix) searches under default parameters for similarityto sequences. The polynucleotide sequence for IPD113Aa (SEQ ID NO: 1)was analyzed. Gene identities conducted by BLAST™ in a DUPONT PIONEERinternal plant transcriptomes database identified multiple homologs ofIPD113Aa protein (SEQ ID NO: 1). The IPD113Aa homologs and the organismthey were identified from are shown in Table 3.

TABLE 3 Gene Name Source Organism DNA Seq AA Seq IPD113Aa PS930 Pterisalbolineata syn Pteris cretica cv albolineata SEQ ID NO: 127 SEQ ID NO:1 IPD113Ab PS930 Pteris albolineata syn Pteris cretica cv albolineataSEQ ID NO: 128 SEQ ID NO: 2 IPD113Ac PS930 Pteris albolineata syn Pteriscretica cv albolineata SEQ ID NO: 129 SEQ ID NO: 3 IPD113Ad PS930 Pterisalbolineata syn Pteris cretica cv albolineata SEQ ID NO: 130 SEQ ID NO:4 IPD113Ae PS995 Pteris umbrosa SEQ ID NO: 131 SEQ ID NO: 5 IPD113BaPS12357 Polypodium formosanum ‘Cristatum’ SEQ ID NO: 132 SEQ ID NO: 6IPD113Bb PS12357 Polypodium formosanum ‘Cristatum’ SEQ ID NO: 133 SEQ IDNO: 7 IPD113Bc PS12357 Polypodium formosanum ‘Cristatum’ SEQ ID NO: 134SEQ ID NO: 8 IPD113Da PS8824 Nephrolepis obliterata ‘Kimberly Queen’ SEQID NO: 135 SEQ ID NO: 9 IPD113Db PS7897 Colysis wrightii (Hook.) ChingSEQ ID NO: 136 SEQ ID NO: 10 IPD113Dc PS8847 Nephrolepis exaltata‘Compacta’ SEQ ID NO: 137 SEQ ID NO: 11 IPD113Dd PS12356 Davalliatyermannii (orig: Humata tyermannii) SEQ ID NO: 138 SEQ ID NO: 12IPD113De PS14958 Pyrrosia lanceolata SEQ ID NO: 139 SEQ ID NO: 13IPD113Df PS14958 Pyrrosia lanceolata SEQ ID NO: 140 SEQ ID NO: 14IPD113Dg PS14958 Pyrrosia lanceolata SEQ ID NO: 141 SEQ ID NO: 15IPD113Dh PS9539 Tectaria milnei SEQ ID NO: 142 SEQ ID NO: 16 IPD113DiPS2138 Polystichum proliferum SEQ ID NO: 143 SEQ ID NO: 17 IPD113DjPS2138 Polystichum proliferum SEQ ID NO: 144 SEQ ID NO: 18 IPD113DkPS2138 Polystichum proliferum SEQ ID NO: 145 SEQ ID NO: 19 IPD113DlPS13705 Polystichum acrostichoides SEQ ID NO: 146 SEQ ID NO: 20 IPD113DmPS845 Pyrrosia rupestris SEQ ID NO: 147 SEQ ID NO: 21 IPD113Dn PS845Pyrrosia rupestris SEQ ID NO: 148 SEQ ID NO: 22 IPD113Do PS9163Asplenium antiquum Makino SEQ ID NO: 149 SEQ ID NO: 23 IPD113Dp NY28Doryopteris cordata SEQ ID NO: 150 SEQ ID NO: 24 IPD113Dq NY26 Aspleniumebenoides SEQ ID NO: 151 SEQ ID NO: 25 IPD113Dr NY26 Asplenium ebenoidesSEQ ID NO: 152 SEQ ID NO: 26 IPD113Ds NY100 Adiantum venustum SEQ ID NO:153 SEQ ID NO: 27 IPD113Ds NY100 Adiantum venustum SEQ ID NO: 154 SEQ IDNO: 28 (M18 Start) IPD113Dt NY75 Arachniodes standishii SEQ ID NO: 155SEQ ID NO: 29 IPD113Du NY100 Adiantum venustum SEQ ID NO: 156 SEQ ID NO:30 IPD113Ea NY007 Blechnum medium (originally Doodia media) SEQ ID NO:157 SEQ ID NO: 31 IPD113Eb PS12888 Dryopteris intermedia SEQ ID NO: 158SEQ ID NO: 32 IPD113Ec PS12888 Dryopteris intermedia SEQ ID NO: 159 SEQID NO: 33 IPD113Ed PS12888 Dryopteris intermedia SEQ ID NO: 160 SEQ IDNO: 34 IPD113Ee PS9145 Ophioglossum pendulum SEQ ID NO: 161 SEQ ID NO:35 IPD113Ef PS9145 Ophioglossum pendulum SEQ ID NO: 162 SEQ ID NO: 36IPD113Eg PS9145 Ophioglossum pendulum SEQ ID NO: 163 SEQ ID NO: 37IPD113Eh PS14994 Pellaea falcata SEQ ID NO: 164 SEQ ID NO: 38 IPD113EiPS989 Adiantum aethiopicum SEQ ID NO: 165 SEQ ID NO: 39 IPD113Ej PS989Adiantum aethiopicum SEQ ID NO: 166 SEQ ID NO: 40 IPD113Fa PS9224Lygodium flexuosum SEQ ID NO: 167 SEQ ID NO: 41 IPD113Fb PS9224 Lygodiumflexuosum SEQ ID NO: 168 SEQ ID NO: 42 IPD113Fc PS9224 Lygodiumflexuosum SEQ ID NO: 169 SEQ ID NO: 43 IPD113Fd PS9224 Lygodiumflexuosum SEQ ID NO: 170 SEQ ID NO: 44 IPD113Fe PS930 Pteris albolineatasyn Pteris cretica cv albolineata SEQ ID NO: 171 SEQ ID NO: 45 IPD113FfPS930 Pteris albolineata syn Pteris cretica cv albolineata SEQ ID NO:172 SEQ ID NO: 46 IPD113Fg PS930 Pteris albolineata syn Pteris creticacv albolineata SEQ ID NO: 173 SEQ ID NO: 47 IPD113Fh PS930 Pterisalbolineata syn Pteris cretica cv albolineata SEQ ID NO: 174 SEQ ID NO:48 IPD113Fi NY14 Adiantum polyphyllum ‘Amaretto’ SEQ ID NO: 175 SEQ IDNO: 49 IPD113Fj NY14 Adiantum polyphyllum ‘Amaretto’ SEQ ID NO: 176 SEQID NO: 50 IPD113Fk NY14 Adiantum polyphyllum ‘Amaretto’ SEQ ID NO: 177SEQ ID NO: 51 IPD113Fl PS8798 Blechnum occidentale SEQ ID NO: 178 SEQ IDNO: 52 IPD113Ga NY007 Blechnum medium (originally Doodia media) SEQ IDNO: 179 SEQ ID NO: 53 IPD113Gb NY54 Gymnocarpium dryopteris SEQ ID NO:180 SEQ ID NO: 54 IPD113Gc NY54 Gymnocarpium dryopteris SEQ ID NO: 181SEQ ID NO: 55 IPD113Gd NY54 Gymnocarpium dryopteris SEQ ID NO: 182 SEQID NO: 56 IPD113Ge NY54 Gymnocarpium dryopteris SEQ ID NO: 183 SEQ IDNO: 57 IPD113Gf PS13705 Polystichum acrostichoides SEQ ID NO: 184 SEQ IDNO: 58 IPD113Gg PS13705 Polystichum acrostichoides SEQ ID NO: 185 SEQ IDNO: 59 IPD113Gh PS13705 Polystichum acrostichoides SEQ ID NO: 186 SEQ IDNO: 60 IPD113Gi PS898 Cheilanthes sieberi SEQ ID NO: 187 SEQ ID NO: 61IPD113Dv PS3637 Polypodium vulgare SEQ ID NO: 188 SEQ ID NO: 62 IPD113EkPS3640 Adiantum hispidulum var. whtei SEQ ID NO: 189 SEQ ID NO: 63IPD113El PS3640 Adiantum hispidulum var. whtei SEQ ID NO: 190 SEQ ID NO:64 IPD113Em PS5307 Colysis ampla SEQ ID NO: 191 SEQ ID NO: 65 IPD113EnPS5307 Colysis ampla SEQ ID NO: 192 SEQ ID NO: 66 IPD113Eo PS826Adiantum formosum SEQ ID NO: 193 SEQ ID NO: 67 IPD113Ep PS826 Adiantumformosum SEQ ID NO: 194 SEQ ID NO: 68 IPD113Eq PS826 Adiantum formosumSEQ ID NO: 195 SEQ ID NO: 69 IPD113Dw PS843 Polypodium billardieri SEQID NO: 196 SEQ ID NO: 70 IPD113Dx PS11034 Polystichum braunii SEQ ID NO:197 SEQ ID NO: 71 IPD113Dy PS9433 Lygodium japonicum SEQ ID NO: 198 SEQID NO: 72 IPD113Dz PS9433 Lygodium japonicum SEQ ID NO: 199 SEQ ID NO:73 IPD113Daa PS9433 Lygodium japonicum SEQ ID NO: 200 SEQ ID NO: 74IPD113Dab NY30 Dryopteris lepidopoda SEQ ID NO: 201 SEQ ID NO: 75IPD113Er PS2220 Pteridium esculentum SEQ ID NO: 202 SEQ ID NO: 76IPD113Es PS3642 Pellaea falcata var. nana SEQ ID NO: 203 SEQ ID NO: 77IPD113Gj PS3642 Pellaea falcata var. nana SEQ ID NO: 204 SEQ ID NO: 78IPD113Fm PS4722 Christella dentata SEQ ID NO: 205 SEQ ID NO: 79 IPD113FnPS4722 Christella dentata SEQ ID NO: 206 SEQ ID NO: 80 IPD113Fo PS4722Christella dentata SEQ ID NO: 207 SEQ ID NO: 81 IPD113Dac PS5237Lastreopsis tinarooensis SEQ ID NO: 208 SEQ ID NO: 82 IPD113Dad PS5237Lastreopsis tinarooensis SEQ ID NO: 209 SEQ ID NO: 83 IPD113Fp PS5239Asplenium boltonii SEQ ID NO: 210 SEQ ID NO: 84 IPD113Fq PS5239Asplenium boltonii SEQ ID NO: 211 SEQ ID NO: 85 IPD113Fr PS5256Campyloneurum xalapense SEQ ID NO: 212 SEQ ID NO: 86 IPD113Fs PS5256Campyloneurum xalapense SEQ ID NO: 213 SEQ ID NO: 87 IPD113Dae PS5307Colysis ampla SEQ ID NO: 214 SEQ ID NO: 88 IPD113Daf PS5307 Colysisampla SEQ ID NO: 215 SEQ ID NO: 89 IPD113Dag PS5307 Colysis ampla SEQ IDNO: 216 SEQ ID NO: 90 IPD113Dah PS5307 Colysis ampla SEQ ID NO: 217 SEQID NO: 91 IPD113Et PS9169 Asplenium dimorphum × difforme SEQ ID NO: 218SEQ ID NO: 92 IPD113Eu NY28 Doryopteris cordata SEQ ID NO: 219 SEQ IDNO: 93 IPD113Ev NY28 Doryopteris cordata SEQ ID NO: 220 SEQ ID NO: 94IPD113Ew NY28 Doryopteris cordata SEQ ID NO: 221 SEQ ID NO: 95 IPD113ExNY28 Doryopteris cordata SEQ ID NO: 222 SEQ ID NO: 96 IPD113Dai PS13456Nephrolepis exaltata SEQ ID NO: 223 SEQ ID NO: 97 IPD113Daj PS13456Nephrolepis exaltata SEQ ID NO: 224 SEQ ID NO: 98 IPD113Dc_M28 PS12360Nephrolepis exaltata ‘Tiger Fern’ SEQ ID NO: 225 SEQ ID NO: 99 IPD113DakPS12360 Nephrolepis exaltata ‘Tiger Fern’ SEQ ID NO: 226 SEQ ID NO: 100IPD113Dal PS12360 Nephrolepis exaltata ‘Tiger Fern’ SEQ ID NO: 227 SEQID NO: 101 IPD113Dam PS12360 Nephrolepis exaltata ‘Tiger Fern’ SEQ IDNO: 228 SEQ ID NO: 102 IPD113Ey NY25 Hemionitis arifolia SEQ ID NO: 229SEQ ID NO: 103 IPD113Ez NY25 Hemionitis arifolia SEQ ID NO: 230 SEQ IDNO: 104 IPD113Eaa NY25 Hemionitis arifolia SEQ ID NO: 231 SEQ ID NO: 105IPD113Eab NY25 Hemionitis arifolia SEQ ID NO: 232 SEQ ID NO: 106IPD113Eac NY25 Hemionitis arifolia SEQ ID NO: 233 SEQ ID NO: 107IPD113Ft PS2140 Pteridium esculentum SEQ ID NO: 234 SEQ ID NO: 108IPD113Fv PS2140 Pteridium esculentum SEQ ID NO: 235 SEQ ID NO: 109IPD113Ead PS14994 Pellaea falcata SEQ ID NO: 236 SEQ ID NO: 110IPD113Dan PS14994 Pellaea falcata SEQ ID NO: 237 SEQ ID NO: 111IPD113Dao PS14994 Pellaea falcata SEQ ID NO: 238 SEQ ID NO: 112IPD113Dap PS022 Ophioglossum pendulum (Medium) SEQ ID NO: 239 SEQ ID NO:113 IPD113Daq PS022 Ophioglossum pendulum (Medium) SEQ ID NO: 240 SEQ IDNO: 114 IPD113Gk PS5338 Selliguea feei SEQ ID NO: 241 SEQ ID NO: 115IPD113Gl PS5338 Selliguea feei SEQ ID NO: 242 SEQ ID NO: 116 IPD113GmPS5338 Selliguea feei SEQ ID NO: 243 SEQ ID NO: 117 IPD113Gn PS5338Selliguea feei SEQ ID NO: 244 SEQ ID NO: 118 IPD113Go PS5338 Selligueafeei SEQ ID NO: 245 SEQ ID NO: 119 IPD113Eai NY165 Polypodiumglycyrrhiza SEQ ID NO: 246 SEQ ID NO: 120 IPD113Eae PS5360 Tectariaantioquoiana SEQ ID NO: 247 SEQ ID NO: 121 IPD113Eah PS5360 Tectariaantioquoiana SEQ ID NO: 248 SEQ ID NO: 122 IPD113Eaf PS5360 Tectariaantioquoiana SEQ ID NO: 249 SEQ ID NO: 123 IPD113Eag PS5360 Tectariaantioquoiana SEQ ID NO: 250 SEQ ID NO: 124 IPD113Fx PS11699 Nephrolepiscordifolia (Duffii) SEQ ID NO: 251 SEQ ID NO: 125 IPD113Fw PS11699Nephrolepis cordifolia (Duffii) SEQ ID NO: 252 SEQ ID NO: 126 IPD113CaPS6040 Asplenium pellucidum SEQ ID NO: 335 SEQ ID NO: 416 IPD113CbPS6069 Asplenium laserpitiifolium SEQ ID NO: 336 SEQ ID NO: 417 IPD113BePS6069 Asplenium laserpitiifolium SEQ ID NO: 337 SEQ ID NO: 418 IPD113BdPS6069 Asplenium laserpitiifolium SEQ ID NO: 338 SEQ ID NO: 419 IPD113BfPS6069 Asplenium laserpitiifolium SEQ ID NO: 339 SEQ ID NO: 420IPD113Eaj PS6087 Hymenasplenium unilaterale SEQ ID NO: 340 SEQ ID NO:421 IPD113Dav PS5404 Pellaea ovata SEQ ID NO: 341 SEQ ID NO: 422IPD113Daw PS5404 Pellaea ovata SEQ ID NO: 342 SEQ ID NO: 423 IPD113DaxPS5404 Pellaea ovata SEQ ID NO: 343 SEQ ID NO: 424 IPD113Day PS5404Pellaea ovata SEQ ID NO: 344 SEQ ID NO: 425 IPD113Daz PS5404 Pellaeaovata SEQ ID NO: 345 SEQ ID NO: 426 IPD113Dba PS5404 Pellaea ovata SEQID NO: 346 SEQ ID NO: 427 IPD113Dar PS5404 Pellaea ovata SEQ ID NO: 347SEQ ID NO: 428 IPD113Das PS5404 Pellaea ovata SEQ ID NO: 348 SEQ ID NO:429 IPD113Dat PS5404 Pellaea ovata SEQ ID NO: 349 SEQ ID NO: 430IPD113Dau PS5404 Pellaea ovata SEQ ID NO: 350 SEQ ID NO: 431 IPD113DbbPS5405 Myriopteris myriophylla SEQ ID NO: 351 SEQ ID NO: 432 IPD113DbdPS5405 Myriopteris myriophylla SEQ ID NO: 352 SEQ ID NO: 433 IPD113DbePS5405 Myriopteris myriophylla SEQ ID NO: 353 SEQ ID NO: 434 IPD113DbcPS5405 Myriopteris myriophylla SEQ ID NO: 354 SEQ ID NO: 435 IPD113FacNY182 Pyrrosia linearifolia SEQ ID NO: 355 SEQ ID NO: 436 IPD113FadNY189 Pyrrosia stigmosa SEQ ID NO: 356 SEQ ID NO: 437 IPD113Fae NY189Pyrrosia stigmosa SEQ ID NO: 357 SEQ ID NO: 438 IPD113Eak LW12339Polypodium attenuatum ‘Falax’ SEQ ID NO: 358 SEQ ID NO: 439 IPD113EalLW12339 Polypodium attenuatum ‘Falax’ SEQ ID NO: 359 SEQ ID NO: 440IPD113Eam LW12339 Polypodium attenuatum ‘Falax’ SEQ ID NO: 360 SEQ IDNO: 441 IPD113Gp LW9210 Aglaomorpha ‘Roberts' SEQ ID NO: 361 SEQ ID NO:442 IPD113Faa LW9210 Aglaomorpha ‘Roberts' SEQ ID NO: 362 SEQ ID NO: 443IPD113Fab LW9210 Aglaomorpha ‘Roberts' SEQ ID NO: 363 SEQ ID NO: 444IPD113Gq LW9210 Aglaomorpha ‘Roberts' SEQ ID NO: 364 SEQ ID NO: 445IPD113Fy LW9210 Aglaomorpha ‘Roberts' SEQ ID NO: 365 SEQ ID NO: 446IPD113Fz LW9210 Aglaomorpha ‘Roberts' SEQ ID NO: 366 SEQ ID NO: 447IPD113Dbf LW9539 Tectaria milnei SEQ ID NO: 367 SEQ ID NO: 448 IPD113DbiLW9539 Tectaria milnei SEQ ID NO: 368 SEQ ID NO: 449 IPD113Dbg LW9539Tectaria milnei SEQ ID NO: 369 SEQ ID NO: 450 IPD113Dbh LW9539 Tectariamilnei SEQ ID NO: 370 SEQ ID NO: 451 IPD113Faf NY28 Doryopteris cordataSEQ ID NO: 371 SEQ ID NO: 452 IPD113Fah NY28 Doryopteris cordata SEQ IDNO: 372 SEQ ID NO: 453 IPD113Fai NY28 Doryopteris cordata SEQ ID NO: 373SEQ ID NO: 454 IPD113Faj NY28 Doryopteris cordata SEQ ID NO: 374 SEQ IDNO: 455 IPD113Fak NY28 Doryopteris cordata SEQ ID NO: 375 SEQ ID NO: 456IPD113Fag NY28 Doryopteris cordata SEQ ID NO: 376 SEQ ID NO: 457IPD113Eas PS935 Asplenium flabellifolium SEQ ID NO: 377 SEQ ID NO: 458IPD113Eat PS935 Asplenium flabellifolium SEQ ID NO: 378 SEQ ID NO: 459IPD113Eau PS5226 Davallia pentaphylla SEQ ID NO: 379 SEQ ID NO: 460IPD113Eay PS5226 Davallia pentaphylla SEQ ID NO: 380 SEQ ID NO: 461IPD113Eav PS5226 Davallia pentaphylla SEQ ID NO: 381 SEQ ID NO: 462IPD113Eaw PS5410 Microsorum commutatum SEQ ID NO: 382 SEQ ID NO: 463IPD113Df_C_TR1 PS5428 Pyrrosia rupestris SEQ ID NO: 383 SEQ ID NO: 464IPD113Eap_C_TR1 PS5428 Pyrrosia rupestris SEQ ID NO: 384 SEQ ID NO: 465IPD113Eaq PS5428 Pyrrosia rupestris SEQ ID NO: 385 SEQ ID NO: 466IPD113Eao PS5428 Pyrrosia rupestris SEQ ID NO: 386 SEQ ID NO: 467IPD113Ean PS5428 Pyrrosia rupestris SEQ ID NO: 387 SEQ ID NO: 468IPD113Ear PS5428 Pyrrosia rupestris SEQ ID NO: 388 SEQ ID NO: 469IPD113Eap PS5428 Pyrrosia rupestris SEQ ID NO: 389 SEQ ID NO: 470IPD113Fal PS5428 Pyrrosia rupestris SEQ ID NO: 390 SEQ ID NO: 471IPD113Dbj PS5431 Arthropteris tenella SEQ ID NO: 391 SEQ ID NO: 472IPD113Dbk PS6057 Stenochlaena palustris SEQ ID NO: 392 SEQ ID NO: 473IPD113Dbl PS6057 Stenochlaena palustris SEQ ID NO: 393 SEQ ID NO: 474IPD113Fam LW8833 Elaphoglossum SEQ ID NO: 394 SEQ ID NO: 475 IPD113FanLW8833 Elaphoglossum SEQ ID NO: 395 SEQ ID NO: 476 IPD113Fao LW8833Elaphoglossum SEQ ID NO: 396 SEQ ID NO: 477 IPD113Eax_N_TR1 NY065Myriopteris lanosa ‘Mighty Tidy’ SEQ ID NO: 397 SEQ ID NO: 478 IPD113EaxNY065 Myriopteris lanosa ‘Mighty Tidy’ SEQ ID NO: 398 SEQ ID NO: 479IPD113Eaz NY177 Drynaria sparsisora SEQ ID NO: 399 SEQ ID NO: 480IPD113Eba NY177 Drynaria sparsisora SEQ ID NO: 400 SEQ ID NO: 481IPD113Ebc LW12274 Thelypteris noveboracensis SEQ ID NO: 401 SEQ ID NO:482 IPD113Ebd LW12274 Thelypteris noveboracensis SEQ ID NO: 402 SEQ IDNO: 483 IPD113Ebf LW12274 Thelypteris noveboracensis SEQ ID NO: 403 SEQID NO: 484 IPD113Ebe LW12274 Thelypteris noveboracensis SEQ ID NO: 404SEQ ID NO: 485 IPD113Ebb LW12349 Pteris ensiformis ‘Evergemiensis' SEQID NO: 405 SEQ ID NO: 486 IPD113Dbm LW12415 Polystichum tripteron SEQ IDNO: 406 SEQ ID NO: 487 IPD113Cc PS8002 Asplenium athertonense SEQ ID NO:407 SEQ ID NO: 488 IPD113Ebg PS6088 Lindsaea brachypoda SEQ ID NO: 408SEQ ID NO: 489 IPD113Ebk PS989 Adiantum aethiopicum SEQ ID NO: 409 SEQID NO: 490 lPD113Ebj PS989 Adiantum aethiopicum SEQ ID NO: 410 SEQ IDNO: 491 IPD113Far NY138 Tectaria cicutaria ‘button ball fern’ SEQ ID NO:411 SEQ ID NO: 492 IPD113Faq NY138 Tectaria cicutaria ‘button ball fern’SEQ ID NO: 412 SEQ ID NO: 493 IPD113Fap NY138 Tectaria cicutaria ‘buttonball fern’ SEQ ID NO: 413 SEQ ID NO: 494 IPD113Ebh NY246 Dryopterishondoensis SEQ ID NO: 414 SEQ ID NO: 495 IPD113Ebi NY246 Dryopterishondoensis SEQ ID NO: 415 SEQ ID NO: 496

cDNA was generated from source organisms with identified homologs fromthe internal database by reverse transcription from total RNA. Homologswere PCR amplified from their respective cDNAs using primers designedfor the coding sequences of each homolog and subcloned into a planttransient vector containing the DMMV promoter. Cloned PCR products wereconfirmed by sequencing.

The amino acid sequence identity of the IPD113 homologs was calculatedusing the Needleman-Wunsch algorithm, as implemented in the Needleprogram (EMBOSS tool suite). The percent sequence identities of IPD113homologs within a selected subgroup are shown in Table 4. The percentsequence identities of selected IPD113 homolog subgroups are shown inTable 5. In a likewise manner one skilled in the art can compare thepercent identity of other groupings of IPD113 homologs. Phylogenic treesof selected subgroups of IPD113 homologs are shown in FIGS. 2, 3, and 4.

TABLE 4 IPD113Ab IPD113Ac IPD113Ad IPD113Ae IPD113Ba IPD113Bb IPD113BcIPD113Aa SEQ 96.0 92.4 91.7 95.2 81.8 88.2 88.2 ID NO: 1 IPD113Ab SEQ —94.9 95.6 93.4 80.7 86.9 86.9 ID NO: 2 IPD113Ac SEQ — — 99.2 91.4 82.087.9 87.9 ID NO: 3 IPD113Ad SEQ — — — 91.1 81.6 87.5 87.5 ID NO: 4IPD113Ae SEQ — — — — 82.0 87.9 87.9 ID NO: 5 IPD113Ba SEQ — — — — — 92.892.6 ID NO: 6 IPD113Bb SEQ — — — — — — 99.8 ID NO: 7

TABLE 5 IPD113Ab IPD113Bb IPD113Bc IPD113Db IPD113Dh IPD113Ei IPD113EjIPD113Fa IPD113Fl IPD113Gg IPD113Gh IPD113Aa SEQ 96.0 88.2 88.2 60.663.1 57.0 56.4 50.6 49.3 32.4 35.0 ID NO: 1 IPD113Ab SEQ — 86.9 86.959.9 62.6 56.9 56.3 49.1 49.2 32.2 34.8 ID NO: 2 IPD113Bb SEQ — — 99.860.4 61.4 54.5 54.0 50.4 48.5 32.3 34.6 ID NO: 7 IPD113Bc SEQ — — — 60.461.4 54.5 54.0 50.4 48.5 32.3 34.6 ID NO: 8 IPD113Db SEQ — — — — 91.569.8 69.2 52.6 54.9 34.2 36.9 ID NO: 10 IPD113Dh SEQ — — — — — 70.8 70.353.2 55.7 33.9 36.4 ID NO: 16 IPD113Ei SEQ — — — — — — 99.1 53.5 52.533.6 35.9 ID NO: 39 IPD113Ej SEQ — — — — — — — 53.5 52.0 33.2 35.5 IDNO: 40 IPD113Fa SEQ — — — — — — — — 45.5 32.7 35.2 ID NO: 41 IPD113FlSEQ — — — — — — — — — 33.0 35.1 ID NO: 52 IPD113Gg SEQ — — — — — — — — —— 89.3 ID NO: 59

Example 5—Agrobacterium-Mediated Transient Expression of IPD113 Homologsin Bean

Activity of IPD113 homologs was measured using a bush bean transientexpression system as described in Example 3. The activity spectra fortested IPD113 homologs are summarized in Table 6, where a “++++”indicates an average activity score of <=10% of leaf disc consumed, a“+++” indicates an average activity score of 11-50% leaf disc consumed,a “+” indicates an average activity score of 51-70% leaf disc consumed,a ‘+’ indicates an average activity score of >70% leaf disc consumed,and “ND” indicates not determined.

TABLE 6 SBL FAW CEW ECB VBC IPD113Aa SEQ ID NO: 1 ++++ ++ ++ ++++ ++++IPD113Ab SEQ ID NO: 2 +++ + ++ + ND IPD113Ac SEQ ID NO: 3 + + +++ + NDIPD113Ad SEQ ID NO: 4 ++ + + + ND IPD113Ae SEQ ID NO: 5 + + + + NDIPD113Ba SEQ ID NO: 6 + + + + ND IPD113Bb SEQ ID NO: 7 +++ + +++ ++ +++IPD113Bc SEQ ID NO: 8 ++++ + +++ ++ +++ IPD113Da SEQ ID NO: 9 ++++ +++++++ +++ ++++ IPD113Db SEQ ID NO: 10 ++++ ++++ ++++ +++ ++++ IPD113DcSEQ ID NO: 11 ++++ +++ ++++ +++ ++++ IPD113Dd SEQ ID NO: 12 ++++ ++ ++++++++ ++++ IPD113De SEQ ID NO: 13 +++ ++++ + ++ ++++ IPD113Df SEQ ID NO:14 +++ ++++ + ++ ++++ IPD113Dg SEQ ID NO: 15 +++ ++++ + +++ ++++IPD113Dh SEQ ID NO: 16 ++++ ++++ ++++ ++++ ++++ IPD113Di SEQ ID NO: 17++++ ++++ +++ +++ ++++ IPD113Dj SEQ ID NO: 18 ++++ +++ +++ ++ +++IPD113Dk SEQ ID NO: 19 ++++ +++ +++ ++++ ++++ IPD113Dl SEQ ID NO: 20 ++++ +++ ++ ++++ IPD113Dm SEQ ID NO: 21 ++++ ++++ + +++ ++++ IPD113Dn SEQID NO: 22 ++++ ++++ ++ ++++ ++++ IPD113Do SEQ ID NO: 23 ++ +++ + ++ ++IPD113Dp SEQ ID NO: 24 ++++ ++++ ++++ +++ ++++ IPD113Dq SEQ ID NO: 25 +++++ +++ +++ ++++ IPD113Dr SEQ ID NO: 26 + ++++ +++ ++ ++++ IPD113Ds SEQID NO: 27 +++ +++ ++ ++ ++++ IPD113Ds SEQ ID NO: 28 ++++ ++++ +++ +++++++ (M18 Start) IPD113Dt SEQ ID NO: 29 + + + + + IPD113Du SEQ ID NO: 30+++ ++ ++ +++ ++++ IPD113Ea SEQ ID NO: 31 ++ + + + ND IPD113Eb SEQ IDNO: 32 + + + + ND IPD113Ec SEQ ID NO: 33 + + ++ + ND IPD113Ed SEQ ID NO:34 ++++ + +++ + ++++ IPD113Ee SEQ ID NO: 35 ++++ +++ ++++ +++ ++++IPD113Ef SEQ ID NO: 36 ++ ++ ++ ++ ++++ IPD113Eg SEQ ID NO: 37 ++++ ++++++ +++ ++++ IPD113Eh SEQ ID NO: 38 + ++ ++ + ++++ IPD113Ei SEQ ID NO:39 ++++ ++++ ++++ ++++ ++++ IPD113Ej SEQ ID NO: 40 ++++ ++++ ++++ +++++++ IPD113Fa SEQ ID NO: 41 ++++ + +++ ++ +++ IPD113Fb SEQ ID NO:42 + + + + + IPD113Fc SEQ ID NO: 43 ++ + + + + IPD113Fd SEQ ID NO:44 + + + ++ + IPD113Fe SEQ ID NO: 45 +++ + + + + IPD113Ff SEQ ID NO: 46++ + ++ +++ + IPD113Fg SEQ ID NO: 47 + ++ + ++ + IPD113Fh SEQ ID NO:48 + +++ + + + IPD113Fi SEQ ID NO: 49 + ++ +++ +++ + IPD113Fj SEQ ID NO:50 + ++ + ++ ++ IPD113Fk SEQ ID NO: 51 + + + + + IPD113Fl SEQ ID NO:52 + ++++ + +++ ++ IPD113Ga SEQ ID NO: 53 + + + + ND IPD113Gb SEQ ID NO:54 + ++ + + + IPD113Gc SEQ ID NO: 55 + ++ + ++ + IPD113Gd SEQ ID NO:56 + + + +++ + IPD113Ge SEQ ID NO: 57 + ++ + ++ + IPD113Gf SEQ ID NO:58 + ++ + +++ + IPD113Gg SEQ ID NO: 59 + +++ + +++ ++ IPD113Gh SEQ IDNO: 60 ++ ++ ++ +++ ++ IPD113Gi SEQ ID NO: 61 + + + ++ ++ IPD113Dv SEQID NO: 62 +++ ++ +++ +++ ++++ IPD113Ek SEQ ID NO: 63 + + + + ND IPD113ElSEQ ID NO: 64 + + + +++ ND IPD113Em SEQ ID NO: 65 + ++ + ++ +++ IPD113EnSEQ ID NO: 66 + ++ + +++ ++ IPD113Eo SEQ ID NO: 67 ++ + + ++ + IPD113EpSEQ ID NO: 68 ++ + +++ +++ +++ IPD113Eq SEQ ID NO: 69 + + +++ ++ +++IPD113Dw SEQ ID NO: 70 + ++ +++ +++ ++++ IPD113Dx SEQ ID NO: 71 + + + ++++++ IPD113Dy SEQ ID NO: 72 + + ++ ++ +++ IPD113Dz SEQ ID NO: 73 ++ +++ + +++ IPD113Daa SEQ ID NO: 74 + + +++ +++ +++ IPD113Dab SEQ ID NO:75 + + + ++ ND IPD113Er SEQ ID NO: 76 + + +++ +++ ++++ IPD113Es SEQ IDNO: 77 +++ + +++ +++ +++ IPD113Gj SEQ ID NO: 78 + + + + + IPD113Fm SEQID NO: 79 ++++ + + + + IPD113Fn SEQ ID NO: 80 ++++ ++ + + + IPD113Fo SEQID NO: 81 ++++ ++ + ++ + IPD113Dac SEQ ID NO: 82 + +++ +++ + +++IPD113Dad SEQ ID NO: 83 + + ++ + ND IPD113Fp SEQ ID NO: 84 + + + ++ +IPD113Fq SEQ ID NO: 85 + ++ + + + IPD113Fr SEQ ID NO: 86 + ++ +++ + +IPD113Fs SEQ ID NO: 87 +++ + +++ +++ ++ IPD113Dae SEQ ID NO: 88 + + ++++ +++ IPD113Daf SEQ ID NO: 89 ++ + + +++ +++ IPD113Dag SEQ ID NO: 90++ + + ++ +++ IPD113Dah SEQ ID NO: 91 + + ++ + ND IPD113Et SEQ ID NO:92 + + + ++ ND IPD113Eu SEQ ID NO: 93 ++++ +++ ++++ ++++ ++++ IPD113EvSEQ ID NO: 94 + + + + ND IPD113Ew SEQ ID NO: 95 ++++ ++ ++++ +++ ++++IPD113Ex SEQ ID NO: 96 ++++ +++ ++++ +++ ++++ IPD113Dai SEQ ID NO: 97+++ ++ ++++ +++ ++++ IPD113Daj SEQ ID NO: 98 ++++ +++ ++++ +++ ++++IPD113Dc_M28 SEQ ID NO: 99 ++++ ++++ ++++ ++++ ++++ IPD113Dak SEQ ID NO:100 ++ + +++ + ++++ IPD113Dal SEQ ID NO: 101 +++ +++ +++ + ++++IPD113Dam SEQ ID NO: 102 ++ ++ ++++ + ++++ IPD113Ey SEQ ID NO: 103 ++++++ ++++ +++ ++++ IPD113Ez SEQ ID NO: 104 ++++ ++ ++++ +++ ++++ IPD113EaaSEQ ID NO: 105 ++++ +++ ++++ +++ ++++ IPD113Eab SEQ ID NO: 106 +++ ++++++ ++ ++++ IPD113Eac SEQ ID NO: 107 ++++ ++ ++++ +++ ++++ IPD113Ft SEQID NO: 108 + + +++ + ND IPD113Fv SEQ ID NO: 109 + + ++ + ND IPD113EadSEQ ID NO: 110 + + +++ + ++++ IPD113Dan SEQ ID NO: 111 +++ +++ +++ +++++++ IPD113Dao SEQ ID NO: 112 + + + + + IPD113Dap SEQ ID NO: 113 +++++++ ++++ +++ ++++ IPD113Daq SEQ ID NO: 114 +++ +++ ++++ +++ ++++IPD113Gk SEQ ID NO: 115 + + + + + IPD113Gl SEQ ID NO: 116 + + ++ ++ +IPD113Gm SEQ ID NO: 117 + ++ + + + IPD113Gn SEQ ID NO: 118 + + + + +IPD113Go SEQ ID NO: 119 + + + + + IPD113Eai SEQ ID NO: 120 + + + + NDIPD113Eae SEQ ID NO: 121 + + +++ + ND IPD113Eah SEQ ID NO: 122 + ++ ++ +ND IPD113Eaf SEQ ID NO: 123 + + + + ND IPD113Eag SEQ ID NO: 124 ++ ++++++ ++ ++++ IPD113Fx SEQ ID NO: 125 + + ++ + + IPD113Fw SEQ ID NO:126 + + ++ + + IPD113Ca SEQ ID NO: 416 − + − − − IPD113Cb SEQ ID NO: 417− − − − − IPD113Be SEQ ID NO: 418 − − − − − IPD113Bd SEQ ID NO: 419 − −− + − IPD113Bf SEQ ID NO: 420 − − − + − IPD113Eaj SEQ ID NO: 421 − − − −− IPD113Dav SEQ ID NO: 422 + ++ ++ + ++ IPD113Daw SEQ ID NO: 423 + ++ ++++ ++ IPD113Dax SEQ ID NO: 424 + ++ ++ + +++ IPD113Day SEQ ID NO: 425 ++++ ++ + ++ IPD113Daz SEQ ID NO: 426 + ++ ++ ++ ++ IPD113Dba SEQ ID NO:427 + ++ ++ ++ ++ IPD113Dar SEQ ID NO: 428 ++ +++ ++ ++ ++ IPD113Das SEQID NO: 429 + +++ ++ + ++ IPD113Dat SEQ ID NO: 430 + ++ ++ + ++ IPD113DauSEQ ID NO: 431 ++ ++ +++ ++ ++ IPD113Dbb SEQ ID NO: 432 − − − − ++IPD113Dbd SEQ ID NO: 433 ++ − + − ++ IPD113Dbe SEQ ID NO: 434 − − + − ++IPD113Dbc SEQ ID NO: 435 − − − − ++ IPD113Fac SEQ ID NO: 436 ++ ++ − +++ IPD113Fad SEQ ID NO: 437 − − − − − IPD113Fae SEQ ID NO: 438 − + − − −IPD113Eak SEQ ID NO: 439 − +++ − − ++ IPD113Eal SEQ ID NO: 440 − +++ − −++ IPD113Eam SEQ ID NO: 441 − +++ − − ++ IPD113Gp SEQ ID NO: 442 − − − −− IPD113Faa SEQ ID NO: 443 ++ − − − − IPD113Fab SEQ ID NO: 444 − − − − −IPD113Gq SEQ ID NO: 445 + − − − − IPD113Fy SEQ ID NO: 446 − − − − −IPD113Fz SEQ ID NO: 447 − + − − − IPD113Dbf SEQ ID NO: 448 − + − − −IPD113Dbi SEQ ID NO: 449 − − − − − IPD113Dbg SEQ ID NO: 450 − − − + −IPD113Dbh SEQ ID NO: 451 − − − + − IPD113Faf SEQ ID NO: 452 − − − + −IPD113Fah SEQ ID NO: 453 − + − − − IPD113Fai SEQ ID NO: 454 − − − − −IPD113Faj SEQ ID NO: 455 − − − − − IPD113Fak SEQ ID NO: 456 − − − − −IPD113Fag SEQ ID NO: 457 − − − − − IPD113Eas SEQ ID NO: 458 − − − + −IPD113Eat SEQ ID NO: 459 − − − − − IPD113Eau SEQ ID NO: 460 − − − + −IPD113Eay SEQ ID NO: 461 + + − + − IPD113Eav SEQ ID NO: 462 − − − − −IPD113Eaw SEQ ID NO: 463 + + ++ + ++ IPD113Df_C_TR1 SEQ ID NO: 464 − − −− − IPD113Eap_C_TR1 SEQ ID NO: 465 − − − − − IPD113Eaq SEQ ID NO: 466 −− − + − IPD113Eao SEQ ID NO: 467 − − − − − IPD113Ean SEQ ID NO: 468 − −− − − IPD113Ear SEQ ID NO: 469 − + − − − IPD113Eap SEQ ID NO: 470 − ++ −− ++ IPD113Fal SEQ ID NO: 471 − − − − − IPD113Dbj SEQ ID NO: 472 ++ ++++ + +++ IPD113Dbk SEQ ID NO: 473 − ++ ++ + ++ IPD113Dbl SEQ ID NO: 474− ++ − − − IPD113Fam SEQ ID NO: 475 − + + − − IPD113Fan SEQ ID NO: 476 −− − − − IPD113Fao SEQ ID NO: 477 − − + + − IPD113Eax_N_TR1 SEQ ID NO:478 +++ + +++ ++ +++ IPD113Eax SEQ ID NO: 479 +++ + +++ ++ +++ IPD113EazSEQ ID NO: 480 + ++ ++ ++ ++ IPD113Eba SEQ ID NO: 481 − + ++ + ++IPD113Ebc SEQ ID NO: 482 + − ++ − ++ IPD113Ebd SEQ ID NO: 483 + − ++ − +IPD113Ebf SEQ ID NO: 484 ++ − ++ − ++ IPD113Ebe SEQ ID NO: 485 + − +++ −++ IPD113Ebb SEQ ID NO: 486 − − + + ++ IPD113Dbm SEQ ID NO: 487 − − ++ +++ IPD113Cc SEQ ID NO: 488 − − − − +++ IPD113Ebk SEQ ID NO: 489 ++ + +++− +++ IPD113Ebj SEQ ID NO: 490 ++ ++ ++ − +++ IPD113Far SEQ ID NO: 491− + − − + IPD113Faq SEQ ID NO: 492 − + − − − IPD113Fap SEQ ID NO: 493 −− − − − IPD113Ebh SEQ ID NO: 494 − − − + − IPD113Ebi SEQ ID NO: 495 − +− − − IPD113Ebg SEQ ID NO: 496 − − − − −

Example 6—Lepidoptera Assays with Purified IPD113 Proteins Expressed inE. coli

Selected IPD113 homologs were subcloned from their respective transientexpression vectors into the NdeI/BamHI sites of E. coli expressionvector pET16B containing a N-terminal 10× His tag. pET16B plasmid DNA,containing the respective IPD113 gene insert, was transformed intocompetent C41 E. coli cells for recombinant protein expression. E. colicells were grown overnight at 30° C. with ampicillin selection theninoculated into fresh 2XYT medium (1:50) and further grown at 37° C. toan optical density of about 0.7. At that point cells were chilled theninduced with 1 mM IPTG. Cultures were further grown at 16° C. for 20hours to induce protein expression. The E. coli expressed proteins werepurified by immobilized metal ion chromatography using Ni-NTA agarose(Qiagen™, Germany) according to the manufacturer's protocols. Purifiedfractions were loaded onto Zeba™ Spin desalting columns (ThermoScientific) pre-equilibrated with 1× TBS buffer (25 mM Tris pH8+150 mMNaCl). The eluted protein was run in diet assays to evaluate theinsecticidal protein effects on larvae of corn earworm (CEW)(Helicoverpa zea), European corn borer (ECB) (Ostrinia nubialis), fallarmyworm (FAW) (Spodoptera frugiperda JE Smith), soybean looper (SBL)(Pseudoplusia includens), and velvet bean caterpillar (VBC) (Anticarsiagemmatalis Hübner). Bioassays against the five pest species, wereconducted using a dilution series of purified N-10xHis-IPD113polypeptides incorporated into an agar-based Lepidoptera diet (SouthlandProducts Inc., Lake Village, Ark.) in a 96-well plate format. Fourreplicates were used per sample. Two to five neonate insects were placedinto each well of the treated plate. After four days of incubation at28° C., larvae were scored for mortality or severity of stunting. Thescores were recorded numerically as dead (3), severely stunted (2)(little or no growth but alive and equivalent to a 1^(st) instarlarvae), stunted (1) (growth to second instar but not equivalent tocontrols), or normal (0). The activity of a series of IPD113 homologs issummarized in Table 7, where a “+” denotes seeing at least stunting(average score of 1) at the highest dose tested and a “−” denotes nostunting at the dose tested.

TABLE 7 Top dose Protein (ppm) SBL FAW ECB CEW VBC WCRW IPD113Da592 + + + + NT NT IPD113Da 509 + NT NT NT + − IPD113Db 967 + + + + + NTIPD113Db 942 + NT NT NT + − IPD113Di 521 + + + + + NT IPD113Df1200 + + + + + NT IPD113Dh 633 + + + + + − IPD113Dg 1313 + + + + + NTIPD113Dn 304 + + + + + NT IPD113Ei 240 + + + + + NT IPD113Fh 140 − − − −− NT IPD113Fi 133 − − − − − NT IPD113Gd 483 − − − − − NT IPD113Dp ~30 +− + + + NT IPD113Dr 538 − + + + + NT IPD113Ds ~50 + + + + + NT (M18Start) IPD113Daj 262 + + + + + NT IPD113Dap 650 + + + + + NT IPD113Daq750 + + + + + NT IPD113Dan 767 + − + + + NT IPD113Dc_M28 671 + + + + +NT IPD113Dc 600 + + + + + NT IPD113Eaa ~38 + − + + + NT “NT” denotes nottested;; “SBL” denotes Soybean Looper; “FAW” denotes Fall Armyworm;“ECB” denotes Eastern Corn Borer; “CEW” denotes Corn Earworm; “ECB”denotes Eastern Corn Borer; “VBC” denotes Velvet Bean Caterpillar;“WCRW” denotes Western Corn Rootworm

Example 7—Construction of IPD113 Chimeras and Variants with MultipleAmino Acid Substitutions

To generate active variants with diversified sequences, chimeras betweenIPD113Aa (SEQ ID NO: 1) and IPD113Db (SEQ ID NO: 10) were generated bymulti-PCR fragment overlap assembly. A total of ten chimeras betweenIPD113Aa and IPD113Db were constructed and cloned into a plant transientvector containing the DMMV promoter. Additionally, variants of IPD113Aawith multiple amino acid changes were generated by family shuffling(Chia-Chun J. Chang et al, 1999, Nature Biotechnology 17, 793-797) thepolynucleotide sequences encoding IPD113Aa (SEQ ID NO: 1) and IPD113Db(SEQ ID NO: 10). In this variant library, a codon optimizedpolynucleotide sequence of IPD113Aa (SEQ ID NO: 331) and the nativepolynucleotide sequence of IPD113Db (SEQ ID NO: 136) were used aslibrary parents. Library variants were cloned into a plant transientvector containing the DMMV promoter. Both chimera and family shufflevariants were screened using a bush bean transient expression system asdescribed in Example 3.

Sequence identity of IPD113Aa variants was calculated using theNeedleman-Wunsch algorithm, as implemented in the Needle program (EMBOSStool suite). The percent identity compared to IPD113Aa (SEQ ID NO: 1),variant designation, nucleotide sequences, and amino acid sequences ofthe resulting IPD113Aa variants are summarized in Table 8.

TABLE 8 % Identity to IPD113Aa (SEQ ID NO: 1) Variant PolynucleotidePolypeptide 71.3 IPD113Aa_Db_Chim_01 SEQ ID NO: 282 SEQ ID NO: 253 88.9IPD113Aa_Db_Chim_02 SEQ ID NO: 283 SEQ ID NO: 254 74.7IPD113Aa_Db_Chim_03 SEQ ID NO: 284 SEQ ID NO: 255 85.5IPD113Aa_Db_Chim_04 SEQ ID NO: 285 SEQ ID NO: 256 79.4IPD113Aa_Db_Chim_05 SEQ ID NO: 286 SEQ ID NO: 257 80.8IPD113Aa_Db_Chim_06 SEQ ID NO: 287 SEQ ID NO: 258 90.3IPD113Aa_Db_Chim_07 SEQ ID NO: 288 SEQ ID NO: 259 69.8IPD113Aa_Db_Chim_08 SEQ ID NO: 289 SEQ ID NO: 260 93.1IPD113Aa_Db_Chim_09 SEQ ID NO: 290 SEQ ID NO: 261 67.0IPD113Aa_Db_Chim_10 SEQ ID NO: 291 SEQ ID NO: 262 63.6 XP-113FSlibDb#2SEQ ID NO: 292 SEQ ID NO: 263 73.2 XP-113FSlibDb#3 SEQ ID NO: 293 SEQ IDNO: 264 67.8 XP-113FSlibDb#5 SEQ ID NO: 294 SEQ ID NO: 265 66.2XP-113FSlibDb#6 SEQ ID NO: 295 SEQ ID NO: 266 76.2 XP-113FSlibDb#9 SEQID NO: 296 SEQ ID NO: 267- 65.4 XP-113FSlibDb#11 SEQ ID NO: 297 SEQ IDNO: 268 64.5 XP-113FSlibDb#12 SEQ ID NO: 298 SEQ ID NO: 269 62.6XP-113FSlibDb#13 SEQ ID NO: 299 SEQ ID NO: 270 65.8 XP-113FSlibDb#16 SEQID NO: 300 SEQ ID NO: 271 68.4 XP-113FSlibDb#17 SEQ ID NO: 301 SEQ IDNO: 272 70.4 XP-113FSlibDb#18 SEQ ID NO: 302 SEQ ID NO: 273 66.5XP-113FSlibDb#19 SEQ ID NO: 303 SEQ ID NO: 274 65.6 XP-113FSlibDb#20 SEQID NO: 304 SEQ ID NO: 275 81 XP-113FSlibDb#21 SEQ ID NO: 305 SEQ ID NO:276 66.2 XP-113FSlibDb#24 SEQ ID NO: 306 SEQ ID NO: 277 63.2XP-113FSlibDb#25 SEQ ID NO: 307 SEQ ID NO: 278 62.8 XP-113FSlibDb#26 SEQID NO: 308 SEQ ID NO: 279 62.6 XP-113FSlibDb#29 SEQ ID NO: 309 SEQ IDNO: 280 76.2 XP-113FSlibDb#30 SEQ ID NO: 310 SEQ ID NO: 281

The activity spectra for tested IPD113Aa variants are summarized inTable 9, where a “++++” indicates an average activity score of <=10% ofleaf disc consumed, a “+++” indicates an average activity score of11-50% leaf disc consumed, a “+” indicates an average activity score of51-70% leaf disc consumed, a ‘+’ indicates an average activity scoreof >70% leaf disc consumed, and “ND” indicates not determined.

TABLE 9 IPD113 Variant SBL FAW CEW ECB VBC IPD113Aa_Db_Chim_01 SEQ IDNO: 253 ++++ ++ +++ ND ++++ IPD113Aa_Db_Chim_02 SEQ ID NO: 254 + + ++ND + IPD113Aa_Db_Chim_03 SEQ ID NO: 255 + + + ND + IPD113Aa_Db_Chim_04SEQ ID NO: 256 + + + ND + IPD113Aa_Db_Chim_05 SEQ ID NO: 257 + + + ND +IPD113Aa_Db_Chim_06 SEQ ID NO: 258 + + ++ ND + IPD113Aa_Db_Chim_07 SEQID NO: 259 + + + ND + IPD113Aa_Db_Chim_08 SEQ ID NO: 260 +++ + ++ ND +++IPD113Aa_Db_Chim_09 SEQ ID NO: 261 + + + ND ++ IPD113Aa_Db_Chim_10 SEQID NO: 262 + + + ND + XP-113FSlibDb#2 SEQ ID NO: 263 ++ + ++++ + ++++XP-113FSlibDb#3 SEQ ID NO: 264 + + + + + XP-113FSlibDb#5 SEQ ID NO:265 + + + + + XP-113FSlibDb#6 SEQ ID NO: 266 + + + + + XP-113FSlibDb#9SEQ ID NO: 267- + + + + ++ XP-113FSlibDb#11 SEQ ID NO: 268 +++ + +++ +++++ XP-113FSlibDb#12 SEQ ID NO: 269 + + + + + XP-113FSlibDb#13 SEQ IDNO: 270 +++ + +++ + ++++ XP-113FSlibDb#16 SEQ ID NO: 271 + + ++ + +++XP-113FSlibDb#17 SEQ ID NO: 272 + + +++ + ++++ XP-113FSlibDb#18 SEQ IDNO: 273 + + + + + XP-113FSlibDb#19 SEQ ID NO: 274 + + + + +++XP-113FSlibDb#20 SEQ ID NO: 275 + + ++ + ++ XP-113FSlibDb#21 SEQ ID NO:276 + + + + + XP-113FSlibDb#24 SEQ ID NO: 277 + + ++++ + +++XP-113FSlibDb#25 SEQ ID NO: 278 +++ +++ +++ + ++++ XP-113FSlibDb#26 SEQID NO: 279 + + + + +++ XP-113FSlibDb#29 SEQ ID NO: 280 ++ + ++ + ++++XP-113FSlibDb#30 SEQ ID NO: 281 + + +++ + +

Example 8—Chimeras Between IPD113 Homologs

To generate active variants with diversified sequences, chimeras betweenIPD113 homologs were generated by multi-PCR fragment overlap assembly.Chimeras between selected IPD113 homologs were constructed and clonedinto a plant transient vector containing the DMMV promoter. Chimeravariants were screened using a bush bean transient expression system asdescribed in Example 3.

Sequence identity of chimeras to IPD113Aa was calculated using theNeedleman-Wunsch algorithm, as implemented in the Needle program (EMBOSStool suite). The percent identity compared to IPD113Aa (SEQ ID NO: 1),variant designation, chimera parents, nucleotide sequences, and aminoacid sequences of the resulting IPD113 chimeras are summarized in Table10.

TABLE 10 % Identity to IPD113Aa (SEQ ID NO: 1) Variant Parent 1 Parent 2Polynucleotide Polypeptide 89.9 pAL-1942 IPD113Aa IPD113DH SEQ ID NO:321 SEQ ID NO: 311 81.4 pAL-1943 IPD113Bb IPD113DH SEQ ID NO: 322 SEQ IDNO: 312 60.7 pAL-1944 IPD113Df IPD113DH SEQ ID NO: 323 SEQ ID NO: 31362.4 pAL-1945 IPD113Dr IPD113DH SEQ ID NO: 324 SEQ ID NO: 314 51.3pAL-1946 IPD113Fl IPD113DH SEQ ID NO: 325 SEQ ID NO: 315 73.2 pAL-1998IPD113DH IPD113Aa SEQ ID NO: 326 SEQ ID NO: 316 69.9 pAL-1999 IPD113DHIPD113Bb SEQ ID NO: 327 SEQ ID NO: 317 63.1 pAL-2000 IPD113DH IPD113DfSEQ ID NO: 328 SEQ ID NO: 318 61.8 pAL-2001 IPD113DH IPD113Dr SEQ ID NO:329 SEQ ID NO: 319 61.1 pAL-2002 IPD113DH IPD113Fl SEQ ID NO: 330 SEQ IDNO: 320

The activity spectra for tested IPD113 chimeras are summarized in Table11, where a “++++” indicates an average activity score of <=10% of leafdisc consumed, a “+++” indicates an average activity score of 11-50%leaf disc consumed, a “++” indicates an average activity score of 51-70%leaf disc consumed, a ‘+’ indicates an average activity score of >70%leaf disc consumed, and “ND” indicates not determined.

TABLE 11 SBL FAW CEW ECB VBC pAL-1942 + + ++ + +++ pAL-1943 + + + + +pAL-1944 +++ ++++ +++ +++ ++++ pAL-1945 +++ +++ ++++ +++ ++++ pAL-1946 ++++ + + ++ pAL-1998 + +++ + ++ +++ pAL-1999 + + + + + pAL-2000 +++ +++++++ +++ ++++ pAL-2001 + + + + +++ pAL-2002 + + + + +

Example 9—Vector Constructs for Expression of IPD113 Polypeptides inPlants

For testing in maize an expression vector, VECTOR 1, was constructed toinclude a transgene cassette containing a gene design encoding IPD113Dh(SEQ ID NO: 16), with the MMV ENH:MMV ENH:BYDV promoter (PCT Pub. No.WO2017095698) and maize ADH1 intron 1 linked to the OS-UBI terminator(PCT Pub. No. WO2018102131) and an expression vector, VECTOR 2, wasconstructed to include a transgene cassette containing a gene designencoding IPD113Dh (SEQ ID NO: 16), with the maize ubiquitin promoterlinked to the PINII terminator (US Publication No. 20140130205).

Example 10—Agrobacterium-Mediated Stable Transformation of Maize

For Agrobacterium-mediated maize transformation of insecticidalpolypeptides, the method of Cho was employed (M. J. Cho et al., PlantCell Rep. 33, 1767-1777 (2014)) using PMI with mannose selection.Briefly, immature embryos (IEs) were isolated from maize and infectedwith an Agrobacterium suspension containing vector constructs for theexpression of IPD113. IEs and Agrobacterium were co-cultivated on solidmedium in the dark at 21° C. for 3 days and subsequently transferred toresting medium without selection agent but supplemented withcarbenicillin (ICN, Costa Mesa, Calif., USA) to eliminate Agrobacterium.IEs were transferred to the appropriate resting medium for 10-11 daysbefore transferring to PMI medium containing mannose (Sigma-AldrichCorp, St Louis, Mo., USA) with antibiotic(s). Multiple rounds ofselection were performed until sufficient quantities of tissue wereobtained. Regenerative green tissues were transferred to PHI-XM medium(E. Wu et al., In Vitro Cell. Dev. Biol. Plant 50, 9-18 (2014)) withmannose selection. Shoots were transferred to tubes containing MSBrooting medium for rooting and plantlets transplanted to soil in pots inthe greenhouse.

Example 11—Insect Control Efficacy of Stable Transformed Corn PlantsAgainst a Spectrum of Lepidopteran Insects

Leaf discs were excised from transformed maize plants and tested forinsecticidal activity of IPD113Dh polypeptide (SEQ ID NO: 16) againstthe European Corn Borer (ECB) (Ostrinia nubilalis), Corn Earworm, (CEW)(Helicoverpa zea), and Fall Armyworm (Spodoptera frugiperda). Theconstructs VECTOR 1 and VECTOR 2 for the expression of IPD113Dh (SEQ IDNO: 16) were used to generate transgenic maize events to test forefficacy against feeding damage caused by lepidopteran pests provided byexpression of these polypeptides. FIG. 5 shows the protection from leaffeeding by European Corn Borer (ECB) (Ostrinia nubilalis), Corn Earworm,(CEW) (Helicoverpa zea), and Fall Armyworm (Spodoptera frugiperda) wasconferred by expression of IPD113Dh gene (SEQ ID NO: 142).

Example 12—Transformation and Regeneration of Soybean (Glycine max)

Transgenic soybean lines are generated by the method of particle gunbombardment (Klein et al., Nature (London) 327:70-73 (1987); U.S. Pat.No. 4,945,050) using a BIORAD Biolistic PDS1000/He instrument and eitherplasmid or fragment DNA. The following stock solutions and media areused for transformation and regeneration of soybean plants:

Stock Solutions:

-   Sulfate 100× Stock:

37.0 g MgSO₄.7H₂O, 1.69 g MnSO₄.H₂O, 0.86 g ZnSO₄.7H₂O, 0.0025 gCuSO₄.5H₂O

-   Halides 100× Stock:

30.0 g CaCl₂.2H₂O, 0.083 g KI, 0.0025 g CoCl₂.6H₂O

-   P, B, Mo 100× Stock:    18.5 g KH₂PO₄, 0.62 g H₃B0₃, 0.025 g Na₂MoO₄.2H₂O-   Fe EDTA 100× Stock:    3.724 g Na₂EDTA, 2.784 g FeSO₄.7H₂O-   2,4-D Stock:    -   10 mg/mL Vitamin        -   B5 vitamins, 1000× Stock:            -   100.0 g myo-inositol, 1.0 g nicotinic acid, 1.0 g                pyridoxine HCl, 10 g thiamine.HCL.

Media (Per Liter):

SB199 Solid Medium:

-   -   1 package MS salts (Gibco/BRL—Cat. No. 11117-066), 1 mL B5        vitamins 1000× stock, 30 g Sucrose, 4 ml 2, 4-D (40 mg/L final        concentration), pH 7.0, 2 g Gelrite

SB1 Solid Medium:

-   -   1 package MS salts (Gibco/BRL—Cat. No. 11117-066), 1 mL B5        vitamins 1000× stock, 31.5 g Glucose, 2 mL 2, 4-D (20 mg/L final        concentration), pH 5.7, 8 g TC agar

SB196:

-   -   10 mL of each of the above stock solutions 1-4, 1 mL B5 Vitamin        stock, 0.463 g (NH4)2 SO4, 2.83 g KNO3, 1 mL 2,4 D stock, 1 g        asparagine, 10 g Sucrose, pH 5.7

SB71-4:

-   -   Gamborg's B5 salts, 20 g sucrose, 5 g TC agar, pH 5.7.

SB103:

-   -   1 pk. Murashige & Skoog salts mixture, 1 mL B5 Vitamin stock,        750 mg MgCl2 hexahydrate, 60 g maltose, 2 g gelrite, pH 5.7.

SB166:

-   -   SB103 supplemented with 5 g per liter activated charcoal.

Soybean Embryogenic Suspension Culture Initiation:

Pods with immature seeds from available soybean plants 45-55 days afterplanting are picked, removed from their shells and placed into asterilized magenta box. The soybean seeds are sterilized by shaking themfor 15 min in a 5% Clorox® solution with 1 drop of ivory soap (i.e., 95mL of autoclaved distilled water plus 5 mL Clorox® and 1 drop of soap,mixed well). Seeds are rinsed using 2, 1-liter bottles of steriledistilled water and those less than 3 mm are placed on individualmicroscope slides. The small end of the seed is cut and the cotyledonspressed out of the seed coat. Cotyledons are transferred to platescontaining SB199 medium (25-30 cotyledons per plate) for 2 weeks, thentransferred to SB1 for 2-4 weeks. Plates are wrapped with fiber tape.After this time, secondary embryos are cut and placed into SB196 liquidmedium for 7 days.

Culture Conditions:

Soybean embryogenic suspension cultures (cv. 93Y21) were maintained in50 mL liquid medium SB196 on a rotary shaker, 100-150 rpm, 26° C. on16:8 h day/night photoperiod at light intensity of 80-100 μE/m2/s.Cultures are subcultured every 7-14 days by inoculating up to ½ dimesize quantity of tissue (clumps bulked together) into 50 mL of freshliquid SB196.

Preparation of DNA for Bombardment:

In particle gun bombardment procedures it is possible to use purified 1)entire plasmid DNA; or 2) DNA fragments containing only the recombinantDNA expression cassette(s) of interest. For every seventeen bombardmenttransformations, 85 μL of suspension is prepared containing 1 to 90picograms (pg) of plasmid DNA per base pair of each DNA plasmid. DNAplasmids or fragments are co-precipitated onto gold particles asfollows. The DNAs in suspension are added to 50 μL of a 10-60 mg/mL 0.6μm gold particle suspension and then combined with 50 μL CaCl₂ (2.5 M)and 20 μL spermidine (0.1 M). The mixture is vortexed for 5 sec, spun ina microfuge for 5 sec, and the supernatant removed. The DNA-coatedparticles are then washed once with 150 μL of 100% ethanol, vortexed andspun in a microfuge again, then resuspended in 85 μL of anhydrousethanol. Five μL of the DNA-coated gold particles are then loaded oneach macrocarrier disk.

Tissue Preparation and Bombardment with DNA:

Approximately 100 mg of two-week-old suspension culture is placed in anempty 60 mm×15 mm petri plate and the residual liquid removed from thetissue using a pipette. The tissue is placed about 3.5 inches away fromthe retaining screen and each plate of tissue is bombarded once.Membrane rupture pressure is set at 650 psi and the chamber is evacuatedto −28 inches of Hg. Following bombardment, the tissue from each plateis divided between two flasks, placed back into liquid media, andcultured as described above.

Selection of Transformed Embryos and Plant Regeneration:

After bombardment, tissue from each bombarded plate is divided andplaced into two flasks of SB196 liquid culture maintenance medium perplate of bombarded tissue. Seven days post bombardment, the liquidmedium in each flask is replaced with fresh SB196 culture maintenancemedium supplemented with 100 ng/mL selective agent (selection medium).For selection of transformed soybean cells the selective agent used canbe a sulfonylurea (SU) compound with the chemical name,2-chloro-N-((4-methoxy-6 methy-1,3,5-triazine-2-yl)aminocarbonyl)benzenesulfonamide (common names: DPX-W4189 and Chlorsulfuron).Chlorsulfuron is the active ingredient in the DuPont sulfonylureaherbicide, GLEAN®. The selection medium containing SU is replaced everytwo weeks for 8 weeks. After the 8 week selection period, islands ofgreen, transformed tissue are observed growing from untransformed,necrotic embryogenic clusters. These putative transgenic events areisolated and kept in SB196 liquid medium with SU at 100 ng/mL foranother 5 weeks with media changes every 1-2 weeks to generate new,clonally propagated, transformed embryogenic suspension cultures.Embryos spend a total of around 13 weeks in contact with SU. Suspensioncultures are subcultured and maintained as clusters of immature embryosand also regenerated into whole plants by maturation and germination ofindividual somatic embryos.

Somatic embryos became suitable for germination after four weeks onmaturation medium (1 week on SB166 followed by 3 weeks on SB103). Theyare then removed from the maturation medium and dried in empty petridishes for up to seven days. The dried embryos are then planted inSB71-4 medium where they are allowed to germinate under the same lightand temperature conditions as described above. Germinated embryos aretransferred to potting medium and grown to maturity for seed production.

Example 13—Identification of Amino Acid Positions Affecting the ProteinStability and Function of IPD113

To identify amino acid positions affecting protein structural stabilityand insecticidal function of IPD113, saturation mutagenesis wasperformed on selected positions within IPD113Dap (SEQ ID: 113). Mutantswere generated by site directed mutagenesis. Resulting library variantswere transformed into E. coli cells then picked and cultured in 96-wellplates for protein expression. Cell lysates were generated by B-PER®Protein Extraction Reagent from Thermo Scientific (3747 N Meridian Rd,Rockford, IL USA 61101) and screened for FAW insecticidal activity.

Table 12 summarizes the amino acid substitutions identified at eachmutagenized position of IPD113Dap (SEQ ID: 113) and amino acidsubstitutions allowing retention of insecticidal activity.

TABLE 12 AA Position Identified substitutions Active substitutions R 043G, A, V, L, I, M, W, F, P, S, T, C, Y, Q, E, K, H R 045 A, K A, K K 057G, A, V, L, I, S, C, Y, N, Q, R G, A, V, L, I, S, C, Y, N, Q, R R 064 I,W, S, Y, D, K I, W, S, Y, D, K K 074 G, A, V, L, M, F, P, S, T, C, Y, Q,D, G, V, L, M, F, S, T, C, Q, D, E, R E, R, H K 082 G, M, F, P, S, C, Y,N, E, R, H G, M, F, P, S, C, Y, N, E, R, H L 083 G, A, V, I, W, F, S, T,C, Y, N, D, K, G, A, V, I, W, F, S, T, C, Y, N, D, R R R 088 G, A, V, L,M, W, F, P, S, T, C, Y, N, G, A, V, L, M, W, F, P, S, T, C, Y, N, Q, D,E, H Q, D, E, H E 094 G, A, V, L, I, P, S, T, Q, D, K, R, H G, A, V, L,I, P, S, T, Q, D, K, R, H L 100 G, A, V, W, F, S, T, N, Q, D, R A, V, W,F, S, T, Q, R R 101 G, A, V, L, I, M, W, F, S, T, C, Y, N, G, A, V, L,I, M, W, F, S, T, C, Y, N, Q, E, H Q, E, H R 116 G, M, W, S, T G, M, W,S, T E 124 L, S, R L, S, R K 125 A, Q, E, R A, Q, E, R R 142 G, A, V, L,I, M, W, F, S, T, C, Y, Q, G, A, V, L, I, M, W, F, S, T, C, Y, Q, D, E,K, H D, E, K, H R 144 G, A, V, L, I, M, F, S, T, C, Y, N, Q, G, A, V, L,I, M, F, S, T, C, Y, N, Q, D, E, H D, E, H E 169 A, I A, I R 170 G, A,F, P, Y, K G, A, F, P, Y, K E 175 G, A, V, L, I, M, W, F, P, C, Y, N, Q,G, A, V, L, I, M, W, F, P, C, Y, N, Q, D, K, R, H D, K, R, H E 176 G, V,L, M, T G, V, L, M, T K 193 G, A, V, L, S, T, C, Y, N, Q, D G, A, V, L,S, T, C, Y, N, Q, D R 196 G, A, V, L, M, P, S, T, C, Y, N, Q G, A, V, L,M, P, S, T, C, Y, N, Q Y 197 G, A, V, L, M, W, F, P, S, N, D, R, H W, FN 204 G, A, V, I, W, F, S, T, C, Y, E, R G, A, V, I, W, S, T, C, Y, E, RK 207 G, A, V, L, I, W, S, T, Y, N, D, R, H G, A, V, L, I, W, S, T, Y,N, D, R, H R 208 G, V, L, I, F, S, T, C, Y, N, Q, E, K G, V, L, I, F, S,T, C, Y, N, Q, E, K K 210 G, A, V, S, Q, R G, A, V, S, Q, R R 213 V, L,F, Y, H V, L, F, Y, H E 218 G, A, V, L, M, W, F, P, S, T, C, Y, N, G, A,V, L, M, W, F, P, S, T, C, Y, N, Q, K, R Q, K, R R 220 G, A, V, L, S, T,C, Y, N, D, E, K, H G, A, V, L, S, T, C, Y, N, D, E, K, H R 224 G, V, L,M, W, F, S, T, Q, K, H G, V, L, M, W, F, S, T, Q, K, H R 225 V, P, T, KV, P, T, K R 234 D 236 G, A, V, P, S, N, Q G, A, V, P, S, N, Q E 243 G,V, L, F, S, Y, K, R G, V, L, F, S, Y, K, R D 244 C C K 245 G, V, L, F,S, T, C, Y, N, D, E, R, H G, V, L, F, S, T, C, Y, N, D, E, R, H R 258 A,V, L, I, M, W, S, T, C, Y, N, D, K A, V, L, I, M, W, S, T, C, Y, N, D, KE 266 G, A, V, L, M, S, T, Y, N, Q, D, K, R, G, A, V, L, M, S, T, Y, N,Q, D, K, R, H H K 272 G, A, V, L, I, M, W, F, S, T, Y, N G, A, V, L, I,M, W, F, S, T, Y, N R 274 G, A, L, S, C, Y G, A, L, S, C, Y K 295 G, A,V, L, W, P, S, T, C, Q, E, R G, A, V, L, W, P, S, T, C, Q, E, R N 300 G,A, V, L, M, W, P, S, T, C, Y, Q, D, G, A, V, L, M, W, P, S, T, C, Y, D,E, E, R, H R, H A 303 G, V, L, M, W, F, S, C, D, E, K, R G, V, L, M, W,S, C, D, E, K, R R 310 G, A, V, L, I, M, W, F, P, S, T, C, Y, G, A, V,L, I, M, W, F, P, S, T, C, Y, D, E, K, H D, E, K, H L 317 G, V, I, M, W,F, S, T, Y, Q, D, K, R V, I, M, W, F, S, T, D, K, R R 318 K K D 324 G, CG, C W 330 G, A, V, L, I, M, F, S, T, E, K, R, H I, F K 334 R R R 337 A,V, L, W, S, T, Q, H A, V, L, W, S, T, Q, H R 339 G, A, V, L, M, W, F, S,T, C, Y, N, D, G, A, V, L, M, W, F, S, T, C, Y, N, D, E, K, H E, K, H R342 L, T L, T D 344 R 348 G, V, L G, V, L D 349 G, V, F G, V, F R 350 G,A, V, L, I, M, S, Y, N, D, K, H G, A, V, L, I, M, S, Y, N, D, K, H E 353G, A, V, L, M, W, F, P, S, C, Y, D, K, G, A, V, L, M, W, F, P, S, C, Y,D, K, R, H R, H R 357 L, I, S, K L, I, S, K D 363 V V S 379 G, A, V, L,W, P, T, C, Y, Q, D, R A, V, L, W, P, T, C, Y, D, R T 387 G, A, L, I, M,P, S, Q, D G, A, L, I, M, P, S, Q, D S 390 G, A, V, L, M, W, F, C, Q, RG, A, V, L, M, W, F, C, Q, R R 391 G, A, V, L, M, W, F, C, Y, N, D G, A,V, L, M, W, F, C, Y, N, D R 397 G, A, V, L, I, W, F, S, T, C, Y, K, H G,A, V, L, I, W, F, S, T, C, Y, K, H D 398 G, A, V, L, I, M, W, F, P, S,T, C, Y, G, A, V, L, I, M, W, F, P, S, T, C, Y, E, K, R, H E, K, R, H R399 G, A, V, L, I, M, W, F, S, T, Y, D, K, G, A, V, L, I, M, W, F, S, T,Y, D, K, H H E 403 G, L, T, C, K, R G, L, T, C, K, R R 405 G, L, Y G, L,Y E 407 G, W G, W D 411 G, C G, C E 421 G, A, V, L, I, M, F, P, S, T, C,Y, Q, G, A, V, L, I, M, F, P, S, T, C, Y, Q, D, K, R D, K, R D 422 G, L,F, S, C, Y, N, R G, L, F, S, C, Y, N, R R 427 L, F, Q L, F, Q K 428 M,F, H M, F, H E 432 G, A, V, L, I, M, W, F, P, S, T, Y, Q, G, A, V, L, I,M, W, F, P, S, T, Y, Q, D, K, R D, K, R R 438 I I R 449 G, A, L, M, W,F, S, T, C, Q, D, E, K G, A, L, M, W, F, S, T, C, Q, D, E, K R 456 G, A,V, L, M, F, S, C, N, D, E, K G, A, V, L, M, F, S, C, N, D, E, K E 457 G,V, L, I, W, F, S, C, Y, N, D, K, R G, V, L, I, W, F, S, C, Y, N, D, K, RD 458 G, A, V, L, I, M, F, P, S, T, Y, N, Q, G, A, V, L, I, M, F, P, S,T, Y, N, Q, E, R, H E, R, H R 463 A, V, L, W, F, P, S, T, C, Y, N, D, E,A, V, L, W, F, P, S, T, C, Y, N, D, E, H H R 471 G, V, L, M, W, F, S, C,N, E G, V, L, M, W, F, S, C, N, E I 484 G, A, V, L, M, W, F, S, C, Y, Q,D, E, G, A, V, L, M, W, F, S, Y, Q, E, K, R K, R

The above description of various illustrated embodiments of thedisclosure is not intended to be exhaustive or to limit the scope to theprecise form disclosed. While specific embodiments of and examples aredescribed herein for illustrative purposes, various equivalentmodifications are possible within the scope of the disclosure, as thoseskilled in the relevant art will recognize. The teachings providedherein can be applied to other purposes, other than the examplesdescribed above. Numerous modifications and variations are possible inlight of the above teachings and, therefore, are within the scope of theappended claims.

These and other changes may be made in light of the above detaileddescription. In general, in the following claims, the terms used shouldnot be construed to limit the scope to the specific embodimentsdisclosed in the specification and the claims.

The entire disclosure of each document cited (including patents, patentapplications, journal articles, abstracts, manuals, books or otherdisclosures) in the Background, Detailed Description, and Examples isherein incorporated by reference in their entireties.

Efforts have been made to ensure accuracy with respect to the numbersused (e.g. amounts, temperature, concentrations, etc.) but someexperimental errors and deviations should be allowed for. Unlessotherwise indicated, parts are parts by weight, molecular weight isaverage molecular weight; temperature is in degrees centigrade; andpressure is at or near atmospheric.

That which is claimed is:
 1. A recombinant insecticidal polypeptidecomprising an amino acid sequence having at least 80% sequence identityto a polypeptide selected from the group consisting of SEQ ID NO: 1, SEQID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97,SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO:102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQID NO: 125, SEQ ID NO: 126, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO:420, SEQ ID NO: 422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQID NO: 426, SEQ ID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO:430, SEQ ID NO: 431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQID NO: 435, SEQ ID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO:440, SEQ ID NO: 441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQID NO: 448, SEQ ID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO:453, SEQ ID NO: 458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQID NO: 466, SEQ ID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO:473, SEQ ID NO: 474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQID NO: 479, SEQ ID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO:483, SEQ ID NO: 484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQID NO: 488, SEQ ID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO:492, SEQ ID NO: 494, and SEQ ID NO: 495, or an amino acid fragment ofany one of said amino acid sequences that retains the activity of theparent amino acid sequence, and a heterologous signal sequence, ahistidine tag, or heterologous transit sequence operably linked to saidamino acid sequence or amino acid fragment thereof.
 2. The insecticidalpolypeptide of claim 1, wherein the insecticidal polypeptide isinsecticidal against a Lepidopteran agricultural pest.
 3. Theinsecticidal polypeptide of claim 1, wherein said heterologous signalsequence, a histidine tag, or heterologous transit sequence is operablylinked to the N-terminus of said amino acid sequence or said amino acidfragment.
 4. A chimeric insecticidal protein comprising: i) a firstpeptide comprising an amino acid of claim 1, or fragment thereof; ii) asecond peptide comprising an amino acid of claim 1, or fragment thereof,wherein the first and second peptides are different and said first andsecond peptides are linked to one another via a covalent bond; and iii)a heterologous signal sequence, a histidine tag, or heterologous transitsequence operably linked to said first or second peptide.
 5. Thechimeric insecticidal protein of claim 4, wherein the chimericinsecticidal protein has altered insect specificity and/or broadeninsecticidal spectrum compared to a polypeptide comprising only a singleamino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8,SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO:13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ IDNO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27,SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO:32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ IDNO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46,SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO:51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ IDNO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65,SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO:70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ IDNO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84,SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO:89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ IDNO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO:103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO:112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO:121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQID NO: 126, SEQ ID NO: 416, SEQ ID NO: 419, SEQ ID NO: 420, SEQ ID NO:422, SEQ ID NO: 423, SEQ ID NO: 424, SEQ ID NO: 425, SEQ ID NO: 426, SEQID NO: 427, SEQ ID NO: 428, SEQ ID NO: 429, SEQ ID NO: 430, SEQ ID NO:431, SEQ ID NO: 432, SEQ ID NO: 433, SEQ ID NO: 434, SEQ ID NO: 435, SEQID NO: 436, SEQ ID NO: 438, SEQ ID NO: 439, SEQ ID NO: 440, SEQ ID NO:441, SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447, SEQ ID NO: 448, SEQID NO: 450, SEQ ID NO: 451, SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO:458, SEQ ID NO: 460, SEQ ID NO: 461, SEQ ID NO: 463, SEQ ID NO: 466, SEQID NO: 469, SEQ ID NO: 470, SEQ ID NO: 472, SEQ ID NO: 473, SEQ ID NO:474, SEQ ID NO: 475, SEQ ID NO: 477, SEQ ID NO: 478, SEQ ID NO: 479, SEQID NO: 480, SEQ ID NO: 481, SEQ ID NO: 482, SEQ ID NO: 483, SEQ ID NO:484, SEQ ID NO: 485, SEQ ID NO: 486, SEQ ID NO: 487, SEQ ID NO: 488, SEQID NO: 489, SEQ ID NO: 490, SEQ ID NO: 491, SEQ ID NO: 492, SEQ ID NO:494, and SEQ ID NO:
 495. 6. An agricultural composition comprising atleast one insecticidal polypeptide of claim
 1. 7. A recombinantpolynucleotide encoding the insecticidal polypeptide of claim
 1. 8. Therecombinant polynucleotide of claim 7, wherein the polynucleotide isoperably linked to a heterologous regulatory element.
 9. The recombinantpolynucleotide of claim 7, wherein the polynucleotide has codonsoptimized for expression in an agriculturally important crop.
 10. Therecombinant polynucleotide of claim 7, wherein the polynucleotide is acDNA.
 11. A recombinant polynucleotide encoding the chimericinsecticidal protein of claim
 4. 12. A DNA construct comprising therecombinant polynucleotide of claim 7 wherein the DNA construct furthercomprises a gene silencing element or a polynucleotide encoding saidinsecticidal polypeptide, wherein said gene silencing element orinsecticidal polypeptide introduces a trait selected from the groupconsisting of herbicide resistance and disease resistance.
 13. Atransgenic plant comprising the recombinant polynucleotide of claim 7.14. A transgenic plant comprising the DNA construct of claim
 12. 15. Amethod of inhibiting growth or killing an insect pest or pest populationcomprising contacting the insect pest with the insecticidal polypeptideof claim
 1. 16. A method of controlling insect pest damage to plantscomprising providing the insecticidal polypeptide of claim 1 to saidinsect pest or pest population for ingestion, wherein said insecticidalpolypeptide is produced by a transgenic plant and is present in at leastone of said plants.
 17. A method of inhibiting growth or killing aninsect pest or pest population comprising expressing in a transgenicplant the polynucleotide of claim 7, and allowing said insect pest tofeed on said transgenic plant.
 18. A method for controlling an insectpest infestation comprising providing in the diet of the pest thetransgenic plant of claim 16 or a part thereof.
 19. A method forimproving the yield of a crop growing in the presence of an insect pest,said method comprising growing the transgenic plant of claim 14 at leastas part of the crop, wherein the yield of a crop exposed to the insectpest and comprising said transgenic plant has an increased yieldrelative to a crop exposed to the insect pest and not comprising saidtransgenic plant.
 20. The method of claim 19 wherein the ratio saidtransgenic plants to plants lacking said recombinant polynucleotide isabout 1:5, 1:4, 1:1 or 2:1.
 21. The method of claim 17, wherein thetransgenic plant is selected from corn, soybean, wheat, rice, sorghum,sunflower, canola, barley, sugarcane, potatoes, tomatoes, cotton, rapeseed, peanut, and alfalfa.
 22. The method of claim 17, wherein theinsect pest or insect pest population is an agriculturally importantspecies in the Order Lepidopteran.
 23. The method of claim 23, whereinthe insect pest or insect pest population is corn earworm, European cornborer, fall armyworm, soybean looper, and velvet bean caterpillar. 24.The method of claim 17, wherein the insect pest or pest population isresistant to at least one Bt toxin.